12 1. Amino Acids Study Questions Choose the ONE correct answer. 1.1 The letters A through E designate certain regions on the titration curve for glycine (shown below). Which one of the following statements concerning this curve is correct? 2 = nN > ce] = Equivalents OH” added o a A. Point A represents the region where glycine is deprotonated. B. Point B represents a region of minimal buffering.C. Point C represents the region where the net charge on glycine is zero. D. Point D represents the pK of glycine’s carboxyl group. E, Point E represents the pl for glycine. 1.2 Which one of the following statements concerning the peptide shown below is correct? Gly-Cys-Glu-Ser-Asp-Arg-Cys A. The peptide contains glutamine. B. The peptide contains a side chain with a secondary amino group. C. The peptide contains a majority of amino acids with side chains that would be positively charged at pH 7. D. The peptide is able to form an internal disulfide bond. 1.3 Given that the pl for glycine is 6.1, to which electrode, positive or negative, will glycine move in an electric field at pH 2? Explain.Correct answer = C. C represents the isoelectric point or pl, and as such is midway between pK, and pKe for this monoamino monocarboxylic acid. Glycine is fully protonated at Point A. Point B represents a region of maximum buffering, as does Point D. Point E represents the region where glycine is fully deprotonated. NX A ee ry Correct answer = D. The two cysteine residues can, under oxidizing conditions, form a disulfide bond. Glutamine’s 3-letter abbreviation is Gin. Proline (Pro) contains a secondary amino group. Only one (Arg) of the seven would have a posi- tively charged side chain at pH 7.Correct answer = negative electrode. When the PH is less than the pl, the charge on glycine is positive because the o-amino group is fully pro- tonated. (Recall that glycine has H as its R group).24 2, Structure of Proteins Study Questions Choose the ONE correct answer. 2.1 A peptide bond: A. has a partial double-bond character. B. is ionized at physiologic pH. C. is cleaved by agents that denature proteins, such as organic solvents and high concentrations of urea. D. is stable to heating in strong acids. E. occurs most commonly in the cis configuration. 2.2 Which one of the following statements is correct? A. The o-helix can be composed of more than one polypeptide chain. B. B-Sheets exist only in the antiparallel form. C. B-Bends often contain proline. D. Domains are a type of secondary structure. E. The a-helix is stabilized primarily by ionic interac- tions between the side chains of amino acids. 2.3 Which one of the following statements about protein structure is correct?A. Proteins consisting of one polypeptide can have quaternary structure. B. The formation of a disulfide bond in a protein requires that the two participating cysteine residues be adjacent to each other in the primary sequence of the protein. C. The stability of quaternary structure in proteins is mainly a result of covalent bonds among the subunits. D. The denaturation of proteins always leads to irre- versible loss of secondary and tertiary structure. E. The information required for the correct folding of a protein is contained in the specific sequence of amino acids along the polypeptide chain. 2.4 An 80-year-old man presented with impairment of higher intellectual function and alterations in mood and behavior. His family reported progressive disorientation and memory loss over the last 6 months. There is no family history of dementia. The patient was tentatively diagnosed with Alzheimer disease. Which one of the following best describes the disease? A. Itis associated with B-amyloid—an abnormal protein with an altered amino acid sequence. B. It results from accumulation of denatured proteins that have random conformations. C. It is associated with the accumulation of amyloid precursor protein.D. It is associated with the deposition of neurotoxic amyloid peptide aggregates. E. It is an environmentally produced disease not influ- enced by the genetics of the individual. F. It is caused by the infectious form of a host-cell protein. Correct answer = A. The peptide bond has a partial double-bond character. Unlike its compo- nents—the a-amino and o-carboxyl groups—the -NH and -C=0O of the peptide bond do not accept or give off protons. The peptide bond is not cleaved by organic solvents or urea, but is labile to strong acids. It is usually in the trans configuration.Correct answer = C. 8-Bends often contain pro- line, which provides a kink. The a-helix differs from the B-sheet in that it always involves the coiling of a single polypeptide chain. The B-sheet occurs in both parallel and antiparallel forms. Domains are elements of tertiary struc- ture. The o-helix is stabilized primarily by hydro- gen bonds between the -C=O and —NH- groups of peptide bonds. Correct answer = E. The correct folding of a pro- tein is guided by specific interactions between the side chains of the amino acid residues of a polypeptide chain. The two cysteine residues that react to form the disulfide bond may be a great distance apart in the primary structure (or on sep-arate polypeptides), but are brought into close proximity by the three-dimensional folding of the polypeptide chain. Denaturation may either be reversible or irreversible. Quaternary structure requires more than one polypeptide chain. These chains associate through noncovalent interactions.Correct answer = D. Alzheimer disease is asso- ciated with long, fibrillar protein assemblies con- sisting of B-pleated sheets found in the brain and elsewhere. The disease is associated with abnormal processing of a normal protein. The accumulated altered protein occurs in a B-pleated sheet configuration that is neurotoxic. The AB amyloid that is deposited in the brain in Alzheimer disease is derived by proteolytic cleavages from the larger amyloid precursor protein—a single transmembrane protein expressed on the cell surface in the brain and other tissues. Most cases of Alzheimer disease are sporadic, although at least 5-10% of cases are familial. Prion diseases, such as Creutzfeldt- Jakob, are caused by the infectious form (PrP**) of a host-cell protein (PrP).40 3. Globular Proteins Study Questions Choose the ONE correct answer. 3.1 Which one of the following statements concerning the hemoglobins is correct? A. Fetal blood has a higher affinity for oxygen than does adult blood because Hb F has a decreased affinity for 2,3-BPG. B. Purified Hb F (stripped of 2,3-BPG) has a higher affin- ity for oxygen than does purified Hb A. C. The globin chain composition of Hb F is ade. D. Hb Aj¢ differs from Hb A by a single, genetically deter- mined amino acid substitution. E. Hb Ap appears early in fetal life. 3.2 Which one of the following statements concerning the ability of acidosis to precipitate a crisis in sickle cell anemia is correct? A. Acidosis decreases the solubility of Hb S. B. Acidosis increases the affinity of hemoglobin for oxygen. C. Acidosis favors the conversion of hemoglobin fromthe taut to the relaxed conformation. D. Acidosis shifts the oxygen dissociation curve to the left. E. Acidosis decreases the ability of 2,3-BPG to bind to hemoglobin. 3.3 Which one of the following statements concerning the binding of oxygen by hemoglobin is correct? A. The Bohr effect results in a lower affinity for oxygen at higher pH values. B. Carbon dioxide increases the oxygen affinity of hemoglobin by binding to the C-terminal groups of the polypeptide chains. C. The oxygen affinity of hemoglobin increases as the percentage saturation increases. D. The hemoglobin tetramer binds four molecules of 2,3- BPG. E. Oxyhemoglobin and deoxyhemoglobin have the same affinity for protons (H*).Correct answer = A. Because 2,3-BPG reduces the affinity of hemoglobin for oxygen, the weaker interaction between 2,3-BPG and Hb F results in a higher oxygen affinity for Hb F rela- tive to Hb A. In contrast, if both Hb A and Hb F are stripped of 2,3-BPG, they have a similar affinity for oxygen. Hb F consists of ogy. Hb Aic is a glycosylated form of Hb A, formed nonen- zymically in red cells. Hb Ag is a minor compo- nent of normal adult hemoglobin, first appearing shortly before birth and rising to adult levels (about 2% of the total hemoglobin) by 6 months of age. Correct answer = A. Hb § is significantly less soluble in the deoxygenated form, compared with oxyhemoglobin S. A decrease in pH (acido- sis) causes the oxygen dissociation curve toshift to the right, indicating a decreased affinity for oxygen. This favors the formation of the deoxy, or taut, form of hemoglobin, and can pre- cipitate a sickle cell crisis. The binding of 2,3- BPG is increased, because it binds only to the deoxy form of hemoglobins. ee (ay Correct answer = C. The binding of oxygen at one heme group increases the oxygen affinity of the remaining heme groups in the same molecule. Carbon dioxide decreases oxygen affinity because it lowers the pH; moreover, binding of carbon dioxide to the N-termini stabi- lizes the taut, deoxy form. Hemoglobin binds one molecule of 2,3-BPG. Deoxyhemoglobin has a greater affinity for protons and, therefore, is a weaker acid. "Seer or eeV. Chapter Summary 41 Study Questions (continued) 3.4 B-Lysine 82 in hemoglobin A is important for the bind- ing of 2,3-BPG. In Hb Helsinki, this amino acid has been replaced by methionine. Which of the following should be true concerning Hb Helsinki? A. It should be stabilized in the T, rather than the R, form. B. It should have increased Oo affinity and, conse- quently, decreased delivery of O> to tissues. G. Its Os dissociation curve should be shifted to the right relative to Hb A. D. It results in anemia.Correct answer is B. Substitution of lysine by methionine decreases the ability of negatively charged phosphate groups in 2,3-BPG to bind the § subunits of hemoglobin. Because 2,3BPG decreases the Os affinity of hemoglobin, a reduction in 2,3-BPG should result in increased Os affinity and decreased delivery of Oo to tis- sues. The R form is the high-oxygen-affinity form of hemoglobin. Increased Os affinity (decreased delivery) results in a left shift in the Op. dissociation curve. Decreased ©» delivery is compensated for by increased RBC production.42 3. Globular Proteins Study Questions (continued) 3.5 A 67-year-old man presented to the emergency depart- ment with a 1 week history of angina and shortness of breath. He complained that his face and extremities had a “blue color.” His medical history included chronic stable angina treated with isosorbide dinitrate and nitroglycerin. Blood obtained for analysis was chocolate-colored. Which one of the following is the most likely diagnosis? A. Sickle cell anemia B. Carboxyhemoglobinemia ©. Methemoglobinemia. D. B-Thalassemia E. Hemoglobin SC diseaseCorrect answer = C. Oxidation of the heme component of hemoglobin to the ferric (Fe**) state forms methemoglobin. This may be caused by the action of certain drugs, such as nitrates. The methemoglobinemias are charac- terized by chocolate cyanosis (a brownish-blue coloration of the skin and mucous membranes), and chocolate-colored blood as a result of the dark-colored methemoglobin. Symptoms are related to tissue hypoxia, and include anxiety, headache, dyspnea. In rare cases, coma and death can occur.52 4. Fibrous Proteins Study Questions Choose the ONE best answer. 4.1 A 30-year-old woman presented with progressive shortness of breath. She denied the use of cigarettes. A family history revealed that her sister had suffered from unexplained lung disease. Which one of the fol- lowing etiologies most likely explains this patient's pul- monary symptoms? A. Deficiency of proline hydroxylase B. Deficiency of a;-antitrypsin C. Deficiency in dietary vitamin C D Decreased elastase activity E. Increased collagenase activity4.2 A seven-month-old child “fell over’ while crawling, and now presents with a swollen leg. At age 1 month, the infant had multi- ple fractures in various states of healing — (right clavicle, right humerus, right radius). )) At age 7 months, the infant has a fracture of a bowed femur, secondary to minor trauma (see x-ray at right). The bones are thin, have few trabecula, and have thin cortices. A careful family history ruled out nonaccidental trauma (child abuse) as a cause of the bone fractures. The child is most likely to have a defect in: A. type | collagen. B. type Ill collagen. C. type IV collagen. D. elastin. E. fibrillin. 4.3 What is the differential basis of the liver and lung pathology seen in cy-AT deficiency?Correct answer = B. o,-Antitrypsin deficiency is a genetic disorder that can cause pulmonary emphysema even in the absence of cigarette use. A deficiency of o,-antitrypsin permits increased elastase activity to destroy elastin in the alveolar walls, even in nonsmokers. o4-Anti- trypsin deficiency should be suspected when chronic obstructive pulmonary disease (COPD) develops in a patient younger than 45 years who does not have a history of chronic bronchitis or tobacco use, or when multiple family members develop obstructive lung disease at an early age. Choices A, C, and E refer to collagen, not elastin.Correct answer = A. The child most likely has osteogenesis imperfecta. Most cases arise from a defect in the genes encoding type | collagen. Bones in affected patients are thin, osteoporotic, often bowed with a thin cortex and deficient tra- beculae, and extremely prone to fracture. This patient is affected with type |, osteogenesis imperfecta tarda. The disease presents in early infancy with fractures secondary to minor trauma. The disease may be suspected on pre- natal ultrasound through detection of bowing or fractures of long bones. Type II, osteogenesis imperfecta congenita, is more severe, and patients die of pulmonary hypoplasia in utero or during the neonatal period. Defects in type Ill collagen are the most commen cause of Ehlers- Danlos syndrome, characterized by lethal vascu- lar problems and stretchy skin. Type IV collagen forms networks, not fibrils.With o,-AT deficiency, the liver cirrhosis that can result is due to polymerization and retention of a4-AT in the liver, its site of synthesis. The lung pathology is due to this retention-based defi- ciency in o4-AT ( a serin protease inhibitor or ser- pin) such that elastase (a serine protease) is unopposed.68 5, Enzymes Study Questions Choose the ONE correct answer. 5.1 In cases of ethylene glycol poisoning and its charac- teristic metabolic acidosis, treatment involves correc- tion of the acidosis, removal of any remaining ethylene glycol, and administration of an inhibitor of alcohol dehydrogenase (ADH, alcohol:NAD* oxidore- ductase), the enzyme that oxidizes ethylene glycol to the organic acids that cause the acidosis. Ethanol (grain alcohol) frequently is the inhibitor given to treat ethylene glycol poisoning; it works by competitively inhibiting ADH. As a competitive inhibitor, ethanol: A. increases apparent K,, without affecting Vmax. B. decreases apparent K,, without affecting Vinax. C. increases apparent Vinax without affecting Km. D. decreases apparent Vmax without affecting Km. E. decreases both apparent Vinax and apparent Kn. 5.2 ADH requires NAD* for catalytic activity. In the reaction catalyzed by ADH, an alcohol is oxidized to an alde-hyde as NAD* is reduced to NADH and dissociates from the enzyme. The NAD* is functioning as a (an): . apoenzyme. . coenzyme-cosubstrate. . coenzyme-prosthetic group. . cofactor. . heterotropic effector. moowo pr 5.3 A 70-year-old man was admitted to the emergency room with a 12-hour history of chest pain. Serum cre- atine kinase (CK) activity was measured at admission (day 1) and once daily (Figure 5.24). On day 2 after admission, he experienced cardiac arrhythmia, which was terminated by three cycles of electric cardio- conversion, the latter two at maximum energy. [Note: Cardioconversion is performed by placing two pad- dies, 12 cm in diameter, in firm contact with the chest wall and applying a short electric voltage.] Normal car- diac rhythm was reestablished. He had no recurrence of arrhythmia over the next several days. His chest pain subsided and he was released on day 10. Which one of the following is most consistent with the data presented? A. The patient had a myocardial infarction 48 to 64 hours prior to admission.B. The patient had a myocardial infarction on day 2. C. The patient had angina prior to admission. D. The patient had damage to his skeletal muscle on day 2. E. The data do not permit any conclusion concerning myocardial infarction prior to, or after, admission to the hospital. Patient Patient Atadmission 12 hours after (day 1) cardioconversion (day 2} Figure 5.24 Serum creatine kinase levels.Correct answer = A. In the presence of a com- petitive inhibitor, an enzyme appears to have a lower affinity for substrate, but as the substrate level is increased, the observed velocity approaches Vinax. (See panel B of Figures 5.12 and 5.14 to compare effects of competitive and noncompetitive inhibitors.) Correct answer = B. Coenzymes-cosubstrates are small organic molecules that associate tran- siently with an enzyme and leave the enzyme ina changed form. Coenzyme-prosthetic groups are small organic molecules that associate perma- nently with an enzyme and are returned to their original form on the enzyme. Cofactors are metal ions. Heterotropic effectors are not substrates.Correct answer = D. The GK isoenzyme pattern at admission showed elevated MB isozyme, indi- cating that the patient had experienced a myocardial infarction in the previous 12-24 hours. [Note: 48-64 hours after an infarction, the MB isozyme would have returned to normal val- ues.] On day 2, 12 hours after the cardioconver- sions, the MB isozyme had decreased, indicating no further damage to the heart. However, the patient showed an increased MM isozyme after cardioconversion. This suggests damage to muscle, probably a result of the convulsive mus- cle contractions caused by repeated cardiocon- version, Angina is typically the result of transient spasms in the vasculature of the heart, and would not be expected to lead to tissue death that results in elevation in serum creatine kinase.Study Questions Choose the ONE correct answer. 6.1 A muscle biopsy specimen from a patient with a rare disorder, Luft disease, showed abnormally large mito- chondria that contained packed cristae when exam- ined in the electron microscope. Basal ATPase activity of the mitochondria was seven times greater than nor- mal. From these and other data it was concluded that oxidation and phosphorylation were partially uncou- pled. Which of the following statements about this patient is correct? A. The rate of electron transport is abnormally low. B. The proton gradient across the inner mitochondrial membrane is greater than normal. C. ATP levels in the mitochondria are greater than nor- mal. D. Cyanide would not inhibit electron flow. E. The patient shows hypermetabolism and slevated core temperature. 6.2 Explain why and how the malate-aspartate shuttle moves NADH reducing equivalents from the cytosol to the mitochondrial matrix.6.3 CO binds to and inhibits Complex IV of the electron transport chain. What effect, if any, should this respi- ratory inhibitor have on oxidative phosphorylation? Correct answer = E. When phosphorylation is partially uncoupled from electron flow, one would expect a decrease in the proton gradient across the inner mitochondrial membrane and, hence, impaired ATP synthesis. In an attempt to compensate for this defect in energy capture, metabolism and electron flow to oxygen is increased. This hypermetabolism will be accom- panied by elevated body temperature because the energy in fuels is largely wasted, appearing as heat. The electron transport chain will still be inhibited by cyanide.There is no transporter for NADH in the inner mitochondrial membrane. However, NADH can be oxidized to NAD* by the cytoplasmic isozyme of malate dehydrogenase as oxaloac- etate is reduced to malate. The malate is transported across the inner membrane, and the mitochondrial isozyme of malate dehydro- genase oxidizes it to oxaloacetate as mito- chondrial NAD* is reduced to NADH. This NADH can be oxidized by Complex | of the electron transport chain, generating three ATP through the coupled processes of respiration and oxidative phosphorylation. Inhibition of the electron transport chain by res- piratory inhibitors such as CO results in an inability to maintain the proton gradient. Oxidative phosphorylation is therefore inhibited, as are ancillary reactions, because they also require the proton gradient.90 7. Introduction to Carbohydrates Study Question Choose the ONE correct answer. 7.1 Which of the following statements best describes glu- cose? A. It is a ketose and usually exists as a furanose ring in solution. B. Itis a C-4 epimer of galactose. C. It is utilized in biological systems only in the L-iso- meric form. D. Itis produced from dietary starch by the action of o- amylase. E. Homopolysaccharides of glucose, formed by the action of glycosyltransferases, are always branched molecules that contain only B-glycosidic linkages. 7.2 A young black man entered his physician’s office complaining of bloating and diarrhea. His eyes were sunken and the physician noted additional signs of dehydration. The patient’s temperature was normal.He explained that the episode had occurred following a birthday party at which he had participated in an ice cream eating contest. The patient reported prior episodes of a similar nature following ingestion of a significant amount of dairy products. This clinical pic- ture is most probably due to a deficiency in: A. salivary o-amylase. B. isomaltase, C. pancreatic a-amylase. D. sucrase. E. lactase. 7.3 Routine examination of the urine of an asymptomatic pediatric patient showed a positive reaction with Clinitest (a copper reduction method of detecting reduc- ing sugars), but a negative reaction with the glucose oxidase test. Which one of the following sugars is least likely to be present (assuming a single elevated saccharide)? A. Lactose B. Fructose C. Sucrose D. Xylulose E. Galactose 7.4 a-Glucosidase inhibitors such as acarbose and miglitoltaken with meals are used in the treatment of dia- betes. Explain. What effect should these drugs have on the digestion of lactose? Correct answer = B. Glucose and galactose dif- fer only in configuration around carbon 4, and so are C-4 epimers that are interconvertible by the action of an epimerase. Glucose is an aldose sugar that typically exists as a pyranose ring in solution; fructose, however, is a ketose with a furanose ring. The D-isomeric form of carbohy- drates is most typically the form found in biologic systems, in contrast to amino acids. Salivary amylase does not produce monosaccharides. Homopolysaccharides of glucose include branched glycogen in which the glycosidic link- ages are the & form, as well as unbranched cel- lulose that has B linkages.Correct answer = E. The physical symptoms sug- gest a deficiency in an enzyme responsible for carbohydrate degradation. The symptoms observed following the ingestion of dairy products suggest that the patient is deficient in lactase. Correct answer = C. Clinitest is a nonspecific test that produces a change in color if urine is posi- tive for reducing substances, including reducing sugars (glucose, fructose, galactose, xylulose, lactose), amino acids, ascorbic acid, and certain drugs and drug metabolites. Because sucrose is not a reducing sugar, it is not detected by Clinitest. Glucose oxidase method will not detect increased levels of galactose or other sugars in urine. It is therefore important that a copper reduction method be used as a screening test. In those instances when the capper method is posi- tive and the glucose oxidase method is negative, glucosuria is ruled out.a-Glucosidase inhibitors slow the production of glucose from dietary carbohydrates, thereby reducing the post-prandial rise in blood glucose and facilitating better blood glucose control in dia- betics. These drugs have no effect on lactose digestion because the disaccharide lactose con- tains a B-glycosidic bond, not an a.108 8. Glycolysis Study Questions Choose the ONE best answer. 8.1 Which one of the following statements concerning glycolysis is correct? A. The conversion of glucose to lactate requires the presence of oxygen. B. Hexokinase is important in hepatic glucose metab- olism only in the absorptive period following con- sumption of a carbohydrate-containing meal. C. Fructose 2,6-bisphosphate is a potent inhibitor of phosphofructokinase. D. The regulated reactions are also the irreversible reactions. E. The conversion of glucose to lactate yields two ATP and two NADH. 8.2 The reaction catalyzed by phosphofructokinase-1: A. is activated by high concentrations of ATP and citrate.B. uses fructose 1-phosphate as substrate. C. is the rate-limiting reaction of the glycolytic path- way. D. is near equilibrium in most tissues. E. is inhibited by fructose 2,6-bisphosphate. 8.3 Compared with the resting state, vigorously contract- ing skeletal muscle shows: A, an increased conversion of pyruvate to lactate. B. decreased oxidation of pyruvate to COz and water. C. a decreased NADH/NAD* ratio. D. a decreased concentration of AMP. E. decreased levels of fructose 2,6-bisphosphate. 8.4 A 43-year-old man presented with symptoms of weak- ness, fatigue, shortness of breath, and dizziness. His hemoglobin level was less than 7 g/dl (normal for a male being greater than 13.5 g/dl). Red blood cells isolated from the patient showed abnormally low level of lactate production. A deficiency of which one of the following enzymes would be the most likely cause of this patient’s anemia? A. Phosphoglucose isomerase B. Phosphofructokinase C. Pyruvate kinaseD. Hexokinase E. Lactate dehydrogenase Correct answer =D. Hexokinase, phospho- fructokinase, and pyruvate kinase are all irre- versible and are the regulated steps in glycolysis. The conversion of glucose to lactate (anaerobic glycolysis) is a process that does not involve a net oxidation or reduction and, thus, oxygen is not required. Glucokinase (not hexoki- nase) is important in hepatic glucose metab- olism only in the absorptive period following consumption of a carbohydrate-containing meal. Fructose 2,6-bisphosphate is a potent activator (not inhibitor) of phosphofructokinase. The con- version of glucose to lactate yields two ATP but no net production of NADH.Correct answer = C. Phosphofructokinase-1 is the pace-setting enzyme of glycolysis. It is inhib- ited by ATP and citrate, uses fructose 6-phos- phate as substrate, and catalyzes a reaction that is far from equilibrium. The reaction is activated by fructose 2,6-bisphosphate. Correct answer = A. Vigorously contracting mus- cle shows an increased formation of lactate and an increased rate of pyruvate oxidation com- pared with resting skeletal muscle. The levels of AMP and NADH increase, whereas change in the concentration of fructose 2,6-bisphosphate is not a key regulatory factor in skeletal muscle.Correct answer = C. Decreased lactate produc- tion in the erythrocyte indicates a defect in glycol- ysis. Among patients exhibiting genetic defects of glycolytic enzymes, about 95% show a deficiency in pyruvate kinase. Pyruvate kinase deficiency is the second most common cause (after glucose 6- phosphate dehydrogenase deficiency) of enzyme deficiency—related hemolytic anemia.116 9. Tricarboxylic Acid Cycle Study Questions Choose the ONE correct answer. 9.1 The conversion of pyruvate to acetyl CoA and COs: A. is reversible. B. involves the participation of lipoic acid. C. is activated when pyruvate dehydrogenase (PDH, E,) of the pyruvate dehydrogenase complex is phos- phorylated by PDH kinase in the presence of ATP. D. occurs in the cytosol. E. depends on the coenzyme biotin. 9.2 Which one of the following conditions decreases the oxidation of acetyl CoA by the citric acid cycle? A. Alow ATP/ADP ratio B. A low NADH concentration due to rapid oxidation to NAD* through the respiratory chain C. Alow NAD*/NADH ratio D. A high concentration of AMP E. A low GTP/GDP ratio 9.3 The following is the sum of three steps in the citricacid cycle. A + B+ FAD + H20 > C + FADH2 + NADH Choose the lettered answer that corresponds to the missing “A”, “B”, and “C’ in the equation. Reactant A Reactant B Reactant C A. Succinyl CoA GDP Succinate B. Succinate NAD* Oxaloacetate C. Fumarate NAD* Oxaloacetate D. Succinate NAD* Malate E. Fumarate GTP Malate 9.4 A 1-month-old male showed abnormalities of the ner- vous system and lactic acidosis. Enzyme assay for pyruvate dehydrogenase (PDH) activity on extracts of cultured skin fibroblasts showed 5% of normal activity, with a low concentration (1 x 10 mM) of thiamine pyrophosphate (TPP), but 80% of normal activity when the assay contained a high (0.4 mM) concentra- tion of TPP. Which one of the following statements concerning this patient is most correct? A. Elevated levels of lactate and pyruvate in the blood reliably predict the presence of PDH deficiency. B. The patient is expected to show disturbances in fatty acid degradation. C. A diet consisting of high carbohydrate intake would be expected to be beneficial in this patient.D. Alanine concentration in the blood is expected to be less than normal. E. Administration of thiamine is expected to reduce his serum lactate concentration and improve his clinical symptoms. Correct answer = B. Lipoic acid is an intermedi- ate acceptor of the acetyl group formed in the reaction. Pyruvate dehydrogenase complex cat- alyzes an irreversible reaction that is inhibited when the PDH (E1) component is phosphory- lated. The enzyme complex is located in the mitochondrial matrix. Biotin is utilized by car- boxylases. (een R EET) Correct answer = C. A low NAD*/NADH ratio limits the rates of the NAD*-requiring dehydro- genases. A low ATP/ADP or GTP/GDP ratio stimulates the cycle. AMP does not directly affect the cycle.Correct answer = B. Succinate + NAD* + FAD — oxaloacetate + NADH + FADHs Correct answer = E. The patient appears to have a thiamine-responsive PDH deficiency. The enzyme fails to bind thiamine pyrophos- phate at low concentration, but shows significant activity at a high concentration of the coenzyme. This mutation, which affects the Ky of the enzyme for the coenzyme, is present in some, but not all, cases of PDH deficiency. All inborn errors of PDH are associated with elevated lev- els of lactate, pyruvate, and alanine (the transamination product of pyruvate). Patients routinely show neuroanatomic defects, develop- mental delay, and often early death. Elevated lactate and pyruvate are also observed in pyru- vate carboxylase deficiency, another rare defect in pyruvate metabolism. Because PDH is an integral part of carbohydrate metabolism, a diet low in carbohydrates would be expected to blunt the effects of the enzyme deficiency. By con- trast, fatty acid degradation occurs via conver- aN aA Had eesion [0 acetyl VOA Dy p-Oxldalion, @ process Tat does not involve pyruvate as an intermediate. Thus, fatty acid metabolism is not disturbed in this enzyme deficiency.124 10. Gluconeogenesis Study Questions Choose the ONE correct answer. 10.1 The synthesis of glucose from pyruvate by gluconeo- genesis: A. occurs exclusively in the cytosol. B. is inhibited by an elevated level of glucagon. C. requires the participation of biotin. D. involves lactate as an intermediate. E. requires the oxidation/reduction of FAD. 10.2 Which one of the following statements concerning gluconeogenesis is correct? A. It occurs in muscle. B. Itis stimulated by fructose 2,6-bisphosphate, C. Itis inhibited by elevated levels of acetyl CoA. D. It is important in maintaining blood glucose during the normal overnight fast. E. It uses carbon skeletons provided by degradation of fatty acids.10.3 Which one of the following reactions is unique to glu- coneogenesis? A. Lactate — pyruvate B. Phosphoenolpyruvate — pyruvate C. Oxaloacetate — phosphoenolpyruvate D. Glucose 6-phosphate — fructose 6-phosphate E. 1,3-Bis-phosphoglycerate — 3-phosphoglycerate 10.4 The metabolism of ethanol by alcohol dehydroge- nase (ADH) produces NADH. What effect is the change in the NAD*/NADH expected to have on glu- coneogenesis? Explain. 10.5 Given that acetyl CoA cannot be a substrate for glu- coneogenesis, why is its production in fatty acid oxi- dation essential for gluconeogenesis? 10.6 What effect does AMP have on gluconeogenesis and glycolysis? What enzymes are affected?Correct answer = C. Biotin is the coenzyme- prosthetic group of pyruvate carboxylase. The carboxylation of pyruvate occurs in the mito- chondria. Glucagon stimulates gluconeogenesis. Lactate is not an intermediate in the conversion of pyruvate to glucose; however, pyruvate can be produced from lactate. FAD is not involved in gluconeogenesis. Ne (ean een amnnam Correct answer = D. During the overnight fast, glycogen is partially depleted and gluconeo- genesis provides blood glucose. Gluconeo- genesis is inhibited by fructose 2,6-bisphosphate and stimulated by elevated levels of acetyl CoA. Degradation of fatty acids yields acetyl CoA, which cannot be converted to glucose. This is because there is no net gain of carbons from acetyl CoA in the TCA cycle, and the PDH reac- tion is physiologically irreversible. Carbon skele- tons of most amino acids are, however, gluconeogenic.NX Correct answer = C. The other reactions are common to both gluconeogenesis and glycolysis. The increase in NADH as ethanol is oxidized, will decrease the availability of OAA because the reversible oxidation of malate to OAA by malate dehydrogenase of the TCA cycle is driven in the reverse direction by the high avail- ability of NADH. Additionally, the reversible reduction of pyruvate to lactate by lactate dehy- drogenase of glycolysis is driven in the forward direction by NADH. Thus, two important gluco- neogenic substrates, OAA and pyruvate, are decreased as a result of the increase in NADH during ethanol metabolism. This will result in a decrease in gluconeogenesis. Acetyl CoA inhibits pyruvate dehydrogenase and activates pyruvate carboxylase, pushing pyru- vate to gluconeogenesis. 2) A "SEES SuSE EESAMP inhibits gluconeogenesis through inhibition of fructose 1,6-bisphosphatase and favors glycol- ysis through activation of phosphofructokinase-1. (Fructose 2,6-bisohosphate has a similar effect on these enzymes.)136 11. Glycogen Metabolism Study Questions Choose the ONE correct answer. 11.1 A2-year-old boy was brought into the emergency room, suffering from severe fasting hypoglycemia. On physical examination, he was found to have hep- atomegaly. Laboratory tests indicated that he also had hyperlacticacidemia and hyperuricemia. A liver biopsy indicated that hepatocytes contained greater than normal amounts of glycogen that was of normal structure. Enzyme assay likely confirmed a defi- ciency in which of the following enzymes? A. Glycogen synthase B. Glycogen phosphorylase C.Glucose 6-phosphatase D. Amylo-c(1-6)-glucosidase E. Amylo-c:(1>4)->a(1 36)-transglucosidase 11.2 The hormones, epinephrine and glucagon have which one of the following effects on glycogen metabolism in the liver?A. The net synthesis of glycogen is increased. B. Glycogen phosphorylase is phosphorylated and active, whereas glycogen synthase is phosphory- lated and inactive. G. Both glycogen phosphorylase and glycogen syn- thase are activated by phosphorylation but at sig- nificantly different rates. D. Glycogen phosphorylase is inactivated by a rise in Ca?*, whereas glycogen synthase is activated. E. cAMP-dependent protein kinase A is activated, whereas phosphorylase kinase is inactivated. 11.3 In contracting skeletal muscle, a sudden elevation of the cytosolic Ca* concentration will result in: A. activation of cAMP-dependent protein kinase A. B. dissociation of cAMP-dependent protein kinase A into catalytic and regulatory subunits. G. inactivation of phosphorylase kinase caused by the action of protein phosphatase-1. D. direct activation of phosphorylase kinase b. E. direct activation of glycogen phosphorylase b. F. conversion of cAMP to AMP by phosphodiesterase. 11.4 Explain why the hypoglycemia seen with Type la glycogen storage disease (glucose 6-phosphatase deficiency) is severe, whereas that seen with Type VI(liver phosphorylase deficiency) is mild. Correct answer = C. A deficiency of glucose 6- phosphatase (Von Gierke disease) prevents the liver from releasing free glucose into the blood, causing severe fasting hypoglycemia, hyperlacti- cacidemia, and hyperuricemia. A deficiency of glycogen phosphorylase would result in a decrease in glycogen degradation, causing fast- ing hypoglycemia, but not the other symptoms. A deficiency of glycogen synthase would result in lower amounts of stored glycogen. Amylo- a(1-6)-glucosidase removes single glucosyl residues attached to the glycogen chain through an o(1—6)-glycosidic bond. A deficiency in this enzyme would result in a decreased ability of the cell to completely degrade glycogen branches. Amylo-a(1—4)—0(1->6)-transglucosidase defi- ciency would decrease the ability of the cell to make branches.Correct answer = B. Epinephrine and glucagon both cause increased glycogen degradation in the liver through covalent modification (phospho- rylation) of key enzymes of glycogen metabolism. Glycogen phosphorylase is phos- phorylated and active (“a” form), whereas glyco- gen synthase is phosphorylated and inactive (“b” form). cAMP-dependent protein kinase A is active and phosphorylates (and activates) its substrate, phosphorylase kinase. It is phosphory- lase kinase a that directly phosphorylates and activates phosphorylase. Correct answer = D. Ca?* released from the sar- coplasmic reticulum during exercise binds to the calmodulin subunit of phosphorylase kinase, thereby allosterically activating the “b” form of this enzyme. The other choices are not caused by an elevation of cytosolic calcium.With Type la, the liver is unable to generate free glucose either from glycogenolysis or gluconeo- genesis because both processes produce glu- cose 6-phosphate. With Type VI, liver is still able to produce free glucose from gluconeogenesis; glycogenolysis, however, is inhibited.144 12. Metabolism of Monosaccharides and Disaccharides Study Questions Choose the ONE correct answer 12.1 Following the intravenous injection of lactose into a rat, none of the lactose is metabolized. However, ingestion of lactose leads to rapid metabolism of this disaccharide. The difference in these observations is a result of: A. the presence of lactase in the serum. B. the absence of hepatic galactokinase. C.the absence of maltase in the serum. D.the presence of lactase in the intestine. 12.2 A female with classic galactosemia due to GALT defi- ciency is able to produce lactose in breast milk because: A. free (nonphosphorylated) galactose is the accep- tor of glucose transferred by lactose synthase in the synthesis of lactose. B. galactose can be produced from a glucose metabolite by epimerization.C. hexokinase can efficiently phosphorylate dietary galactose to galactose 1-phosphate. D. the enzyme deficient in galactosemia is activated by a hormone produced in the mammary gland. E. galactose can be produced from fructose by iso- merization. 12.3 A 5-month-old boy is brought to his physician because of vomiting, night sweats, and tremors. History revealed that these symptoms began after fruit juices were introduced to his diet as he was being weaned off breast milk. The physical examina- tion was remarkable for hepatomegaly. Tests on the baby’s urine were positive for reducing sugar but negative for glucose. The infant most likely suffers from: A, aldolase B deficiency. B. fructokinase deficiency. C. galactokinase deficiency. D. B-galactosidase deficiency. E. glucose 6-phosphatase deficiency. 12.4 Lactose synthesis is essential in the production of milk by mammary glands. In lactose synthesis: A. galactose from galactose 1-P is transferred to glu- cose by galactosyltransferase (protein A), gener- ating lactose.B. protein A is used exclusively in the synthesis of lactose. C. a-lactalbumin (protein B) regulates the sugar speciticity of protein A by increasing its K, for glu- cose. D. protein B expression is stimulated by prolactin. Correct answer = D. Lactase and maltase are intestinal enzymes not found in the serum. Therefore, ingested lactose is degraded, but injected lactose is not. If hepatic galactokinase is absent, the galactose segment of the lactose is not metabolized, but the glucose segment of the lactose can still be metabolized.Correct answer = B. UDP-hexose 4-epimerase converts UDP-glucose to UDP-galactose, thus providing the appropriate form of galactose for lactose synthesis. UDP-galactose, not free galactose, is the source of the galactose portion of lactose. Galactose is not converted to galac- tose 1-phosphate by hexokinase. Galactosemia is the result of a deficiency in GALT. Isomerization of fructose to galactose does not occur in the human body. Correct answer = A. The symptoms suggest fructose intolerance, a deficiency in aldolase B. Deficiencies in fructokinase or galactokinase result in relatively benign conditions character- ized by elevated levels of fructose or galactose in the blood and urine. Deficiency in B-galactosidase (lactase) results in a decreasedability to degrade lactose (milk sugar). Congenital lactase deficiency is quite rare and would have presented much earlier in this baby, and with different symptoms. Typical lactase deficiency (adult hypolactasia) presents at a later age. The symptoms of glucose 6-phos- phatase deficiency would result from fasting, and would not be related to ingestion of fruit juice. Correct answer = D. The expression of o-lactal- bumin (protein B) is increased by the hormone, prolactin. UDP-galactose is the form used by the galactosyltransferase (protein A). Protein A is also involved in the synthesis of the amino sugar, N-acetyllactosamine. Protein B increases the affinity of protein A for glucose, and so decreases the Km.VI. Chapter Summary 155 Study Questions Choose the ONE correct answer. 13.1 In preparation for a trip to an area of India where malaria is endemic, a young man is given primaquine prophylactically. Soon thereafter, he develops a hemolytic condition. The most likely cause of the hemolysis is a less-than-normal level of which of the following? A. Glucose 6-phosphate B. Oxidized form of NAD C. Reduced form of glutathione D. Ribose 5-phosphate E. Ribulose 5-phosphateCorrect answer = C. Glutathione is essential for red cell integrity and is maintained in its func- tional (reduced) form by NADPH-depencent glu- tathione reductase. The NADPH is generated by the oxidative portion of pentose phosphate path- way. Individuals with a deficiency of the initiating and regulated enzyme (G6PD) of this pathway have a decreased ability to generate NADPH, and thus a decreased ability to keep glutathione functional. When treated with an oxidant drug such as primaquine, some patients with G6PD deficiency develop a hemolytic anemia. Primaquine does not affect glucose 6-phosphate levels. NAD* is neither produced by the pentose phosphate pathway nor used as a coenzyme by glutathione reductase. Ribulose 5-phosphate, another product of the oxidative portion of the pentose phosphate pathway, can be isomerized to ribose 5-phosphate, but a deficiency in either does not cause hemolysis.156 13. Pentose Phosphate Pathway and NADPH 13.2 In male patients who are hemizygous for glucose 6- phosphate dehydrogenase (G6PD) deficiency, patho- physiologic consequences are more apparent in erythrocytes (RBC) than in other cells, such as in the liver. Which one of the following provides the most reasonable explanation for this different response by these individual tissue types? A. Excess glucose 6-phosphate in the liver, but not in RBCs, can be channeled to glycogen, thus avert- ing cellular damage. B. Liver cells, in contrast to RBCs, have alternative mechanisms for supplying the NADPH required for keeping metabolic and cellular integrity. C. Glucose 6-phosphatase activity in RBCs decreases the level of glucose 6-phosphate, thus resulting in cell damage. This does not happen in the hepatocyte. D, Because RBCs do not have mitechondria, produc- tion of ATP required to keep cell integrity depends exclusively on the routing of glucose 6-phosphate to the pentose phosphate pathway. E. The catalytic properties of the liver enzyme are significantly different than those of the RBCenzyme. 13.3.4 G6PD A° individual was treated with primaquine from day 0 to day 120 (Figure 13.15). Hemolysis occurred immediately after initiation of drug therapy, as indicated by progressive anemia, hemoglobinuria, and reticulocytosis. However, despite continued administration of the drug, the hemolysis sponta- neously decreased and red cell survival improved with time. Red cell G6PD activity measured 2 months after termination of therapy was 10% of normal. Which one of the following statements about this patient is correct? A. The patient will continue to be resistant to drug- induced hemolysis after 6 months or longer. B. Erythrocytes in this patient exhibit a longer lifetime than in normal individuals. C. During the period of peak hemolysis, erythrocytes from the patient will show no G6PD activity. D. The intracellular concentration of NADPH in the patient's erythrocytes is greater than normal. E. The patient shows increased bone marrow erythropoiesis.Correct answer = B. Cellular damage is directly related to decreased ability of the cell to regen- erate reduced glutathione, for which large amounts of NADPH are needed, and RBC have no other means of generating NADPH. Catalytic properties of G6PD in liver and RBCs are very similar. The pentose phosphate pathway does not generate ATP. RBCs do not have glucose 6- phosphatase.30 mg Primaquine daily Acute Recovery Resistance or £ 59 hemolysis equilibrium 5 ' ; ‘ 2 ' o ' & ' oO 1 2 ' 2 oO ' = ' a \ se : oc 6 0 8 16) 24 32) 40 114 120 Time (days) Figure 13.15 The course of primaquine-induced hemolysis in a patient with G6PD deficiency.Correct answer = E. As red cells age, the activity of G6PD declines (see Figure 13.13). Despite this loss of enzyme activity, normal old red blood cells contain sufficient G6PD activity to generate NADPH and thereby sustain GSH levels in the face of oxidant stress. In contrast, the G6PD vari- ants with hemolysis have much shorter half-lives. The clinical correlate of this age-related enzyme instability is that hemolysis in patients with G6PD AT generally is mild and limited to the older defi- cient erythrocytes. The anemia is self-limited because the older, vulnerable population of ery- throcytes is replaced by younger RBCs with suffi- cient G6PD activity to withstand an oxidative assault. Although red cell survival remains short- ened as long as use of the drug continues, com- pensation by the erythroid marrow effectively abolishes the anemia in subjects with G6PD A’. The individual's continuing sensitivity to the effects of the drug is revealed by discontinuing the drug for several months to allow the rate of red cell production by the bone marrow to normal- ize; during this phase, the older red cells are able to survive, and the red cell population is rendered sensitive to drug-induced hemolysis. ~172 14. Glycosaminoglycans and Glycoproteins Study Questions Choose the ONE correct answer. 14.1 Mucopolysaccharidoses are inherited lysosomal stor- age diseases. They are caused by: A. an increased rate of synthesis of the carbohydrate component of proteoglycans. B. the synthesis of polysaccharides with an altered structure. CG. defects in the degradation of the GAGs in proteo- glycans. D. the synthesis of abnormally small amounts of pro- tein cores. E. an insufficient amount of proteolytic enzymes. 14.2 The presence of the following compound in the urine of a patient suggests a deficiency in which one of the enzymes listed below? Sulfate Sulfate I |GalNac—GlcUA—GalNAc— A. Galactosidase B. Glucosidase C. Glucuronidase D. Mannosidase E. Sulfatase 14.3 An 8-month-old boy with coarse facial features, skeletal abnormalities, and delays in both growth and development is diagnosed with |-cell disease based on his presentation, and on histologic and biochemi- cal testing. |-cell disease is the consequence of: . decreased production of cell-surface glycopro- teins. . an inability to ubiquitinate proteins. . an inability to glycosylate proteins. . incorrect targeting of lysosomal proteins. increased synthesis of proteoglycans. moog > 14. 4 Distinguish between glycoproteins and proteagly- cans.Correct answer = C. In mucopolysacchari- doses, synthesis of proteoglycans is unaf- fected, both in terms of the structure and the amount of material synthesized. The dis- eases are caused by a deficiency of one of the lysosomal, hydrolytic enzymes responsi- ble for the degradation of glycosaminogly- cans (not the core protein). Correct answer = E. Degradation of glycopro- teins follows the rule “last on, first off.” Because sulfation is the last step in the synthesis of this sequence, a sulfatase is required for the next step in the degradation of the compound shown.Correct answer = D. I-cell disease is a lysosomal storage disease caused by a defect in the gene that initiaties the synthesis of the mannose 6- phosphate signal that targets acid hydrolases to the lysosome, resulting in accumulation of materi- als within the lysosome due to decreased degra- dation. None of the other choices relate in any way to |-cell disease or lysosomal function. Glycoproteins are proteins to which short, branched, oligosaccharide chains are attached. Proteoglycans consist of a core protein to which long, unbranched, glycosaminoglycan (GAG) chains are attached. GAGs are large complexes of negatively charged heteropolysaccharides composed of repeating [acidic sugar-amino sugar]n disaccharide units.180 15. Metabolism of Dietary Lipids Study Questions 15.1Which one of the following statements about the digestion of lipids is correct? A. Lipid digestion begins with acid-stable lipases that primarily use TAG with long to very long-chain fatty acids as substrates. . Large lipid droplets are emulsified (have their sur- face area increased) in the mouth through the act of chewing (mastication). . Colipase facilitates the binding of bile salts to mixed micelles, maximizing the activity of pancreatic lipase. The peptide hormone secretin causes the gallblad- der to contract and release bile. . Patients with cystic fibrosis have difficulties with digestion because their thickened pancreatic secre- tions are less able to reach the small intestine, the primary site of lipid digestion. . Formation of chylomicrons is independent of protein synthesis in the intestinal mucosa. 15.2 Which one of the following statements about theabsorption of lipids from the intestine is correct? A. Dietary triacylglycerol must be completely hydrolyzed to free fatty acids and glycerol before absorption. . The TAG carried by chylomicrons is degraded to free fatty acids and glycerol by lipoprotein lipase on the endothelial surface of capillaries in muscle and adipose primarily. . Fatty acids that contain ten carbons or less are absorbed and enter the circulation primarily via the lymphatic system. . Deticiencies in the ability to absorb fat result in excessive amounts of chylomicrons in the blood.Correct answer = E. Patients with CF, a genetic disease due to a deficiency of functional CFTR, have thickened secretions that impede the flow of pancreatic enzymes into the duodenum. The acid-stable lipases, lingual and gastric lipase, use as substrates TAG with short to medium- chain fatty acids that are abundant in milk. Emulsification occurs through peristalsis that provides mechanical mixing, and bile salts that function as detergents. Colipase restores activity to pancreatic lipase in the presence of inhibitory bile acids that bind the micelles. CCK (cholecys- tokinin) is the hormone that causes contraction of the gallbladder and release of stored bile; secretin causes release of bicarbonate. Chylomicron formation requires synthesis of the protein apolipoprotein B-48. egCorrect answer = B. The TAG in chylomicrons are degraded to fatty acids and glycerol by lipoprotein lipase on the endothelial surface of capillaries in muscle and adipose, thus providing a source of fatty acids to these tissues for degradation or storage. In the duodenum, TAG are degraded to 1 monoacylglycerol + 2 free fatty acids that get absorbed. Medium and short chain fatty acids enter directly into blood; they do not get packaged into chylomicrons. Chylomicrons contain dietary lipids that were digested and absorbed, thus a defect in fat absorption would result in decreased production of chylomicrons. >198 16. Fatty Acid and Triacylglycerol Metabolism Study Questions Choose the ONE correct answer. 16.1 Triacylglycerol molecules stored in adipose tissue represent the major reserve of substrate providing energy during a prolonged fast. During such a fast: A. the stored fatty acids are released from adipose tissue into the plasma as components of the serum lipoprotein particle, VLDL. B. free fatty acids are produced at a high rate in the plasma by the action of lipoprotein lipase on chy- lomicrons. C. glycerol produced by the degradation of triacyl- glycerol is an important direct source of energy for adipocytes and fibroblasts. D. hormone-sensitive lipase is phosphorylated and activated by a cAMP-activated protein kinase.Correct answer = D. Hormone-sensitive lipase is phosphorylated by cAMP-activated protein kinase, which is itself activated by epinephrine or glucagon. Fatty acids released from adipose tissue are carried in the plasma by serum albu- min, not VLDL. During a fast, the amount of cir- culating triacylglycerol (found in chylomicrons and VLDL) will be low. Therefore, there is little substrate for lipoprotein lipase. The glycerol pro- duced during triacylglycerol degradation cannot be metabolized by adipocytes or fibroblasts, but rather must go to the liver where it can be phos- phorylated (by glycerol kinase).200 16. Fatty Acid and Triacylglycerol Metabolism 16.2 A low level of carbon dioxide labeled with '4C is acci- dentally released into the atmosphere surrounding industrial workers as they resume work following the lunch hour. Unknowingly, they breathe the contami- nated air for 1 hour. Which of the following com- pounds will be radioactively labeled? A. All of the carbon atoms of newly synthesized fatty acid. B. About one half of the carbon atoms of newly syn- thesized fatty acids. G. The carboxyl atom of newly synthesized fatty acids. D. About one third of the carbons of newly synthe- sized malonyl CoA. E. One half of the carbon atoms of newly synthe- sized acetyl CoA. 16.3 A teenager, concerned about his weight, attempts to maintain a fat-free diet for a period of several weeks. lf his ability to synthesize various lipids were exam- ined, he would be found to be most deficient in his ability to synthesize:A. triacylglycerol. B. phospholipids. C. cholesterol. D. sphingolipids. E. prostaglandins. 16.4 A 6-month-old boy was hospitalized following a seizure. History revealed that for several days prior, his appetite was decreased due to a “stomach virus’. At admission, his blood glucose was 24 mg/dl (age- referenced normal is 60-100). His urine was negative for ketone bodies, but positive for a variety of dicar- boxylic acids. A tentative diagnosis of medium-chain fatty acyl CoA dehydrogenase (MCAD) deficiency is made. In patients with MCAD deficiency, the fasting hypoglycemia is a consequence of: A. decreased acetyl CoA production. B. decreased ability to convert acetyl CoA to glucose. C. increased conversion of acetyl CoA to acetoac- etate. D. increased production of ATP and NADH. 16.5 Explain why with Zellweger syndrome both very long chain fatty acids (VLCFA) and phytanic acid accumu- late, whereas with X-linked adrenoleukodystrophy (X- ALD) only VLCFA accumulate.Correct answer = D. Malonyl CoA (three car- bons) is synthesized from acetyl CoA (two car- bons) by the addition of CO2, using the enzyme acetyl CoA carboxylase. Because COz is subse- quently removed during fatty acid synthesis, the radioactive label will not appear at any position in newly synthesized fatty acids. Correct answer = E. Prostaglandins are synthe- sized from arachidonic acid. Arachidonic acid is synthesized from linoleic acid, an essential fatty acid obtained by humans from dietary lipids. The teenager would be able to synthesize all other compounds, but presumably in somewhat depressed amounts. "SEES eeCorrect answer = A. Impaired oxidation of fatty acids less than 12 carbons in length results in decreased production of acetyl CoA, the allosteric activator of pyruvate carboxylase, a gluconeogenic enzyme; thus, glucose levels fall. Acetyl CoA cannot be used for the net synthesis of glucose. Acetoacetate is a ketone body, and with MCAD deficiency ketogenesis is decreased. Impaired fatty acid oxidation means that less ATP and NADH are made, and both are needed for gluconeogenesis. Zellweger syndrome is caused by an inability to target matrix proteins to the peroxisome; thus, all peroxisomal activities are affected because functional peroxisomes are not able to be formed. In X-ALD, the defect is an inability to transport VLCFA into the peroxisome—other peroxisomal functions such as a oxidation are normal.218 17. Complex Lipid Metabolism Study Questions Choose the ONE correct answer. 17.1 Aspirin-induced asthma (AIA) is a severe reaction to nonstercidal anti-inflammatory drugs (NSAIDs) char- acterized by bronchoconstriction 30 minutes to sev- eral hours after ingestion. It is seen in as many as 20% of adults. Which of the following statements best explains the symptoms seen in patients with AIA? A. NSAIDs inhibit the activity of the CFTR protein, resulting in thickened secretions that block air- ways. B. NSAIDs inhibit COX but not lipoxygenase, result- ing in the flow of arachidonic acid to leukotriene synthesis. C. NSAIDs activate the COX activity of PGH syn- thase, resulting in increased synthesis of prostaglandins that promote vasodilation. D. NSAIDs activate phospholipases, resulting indecreased amounts of dipalmytoylphosphatidyl- choline and alveolar collapse (atelectasis). 17.2 An infant, born at 28 weeks of gestation, rapidly gave evidence of respiratory distress, Lab and x-ray results supported the diagnosis of infant respiratory distress syndrome (RDS). Which of the following statements about this syndrome is true? A. It is unrelated to the baby’s premature birth. B. It is a consequence of too few Type II pneumo- cytes. C. The lecithin/sphingomyelin ratio in the amniotic fluid is likely to be greater than two. D. The concentration of dipalmitoylphosphatidyl- choline in the amniotic fluid would be expected to be lower than that of a full-term baby. E. RDS is an easily treated disorder with low mortality. 17.3A 25-year-old woman with a history that included hepatosplenomegaly with eventual removal of the spleen, bone and joint pain with several fractures of the femur, and a liver biopsy that showed wrinkled- looking cells with accumulations of glucosylcer- amides was presented at Grand Rounds. The likely diagnosis for this patient is: A. Fabry disease.B. Farber disease. C. Gaucher disease. D. Krabbe disease. E. Niemann-Pick disease. Correct answer = B. NSAIDs inhibit COX but not lipoxygenase, so any arachidonic acid avail- able is used for the synthesis of bronchocon- stricting-leukotrienes. NSAIDs have no effect on the CFTR protein, defects in which are the cause of cystic fibrosis. Steroids, not NSAIDs, inhibit phospholipase As. COX is inhibited by NSAIDs, not activated. NSAIDs have no effect on phospholipases. Correct answer = D. Dipalmitoylphosphatidyl- choline (DPPC, or dipalmitoyl lecithin) is the lung surfactant found in mature, healthy lungs. RDS can occur in lungs that make too little of this compound. If the lecithin/sphingomyelin ratio in amniotic is greater than two, a new-born’s lungs are considered to be sufficiently mature—premature lungs would be expected to have a ratio lower than two. The RDS would not be due to too few Type II pneumocytes—these cells would simply be secreting sphingomyelin rather than DPPC at 28 weeks of gestation. Correct answer = C. The adult form of Gaucher disease causes hepatosplenomegaly, osteo- porosis of the long bones, and the characteristic wrinkled appearance of the cytosol of cells. This is also the sphingolipidosis in which glucosylcer- amides accumulate. The deficient enzyme is B- glucosidase (a glucocerebrosidase).242 18, Cholesterol and Steroid Metabolism Study Questions Choose the ONE correct answer. 18.1 A 95-year-old woman was seen in the emergency room because of racurrent abdominal pain. The his- tory revesled @ 2-year pattern of pain in the upper right quadrant, beginning several hours after the ingestion al a meal rich in friad/latty laod. Ultrasonographic examination demonstrated the presence of numerous stones in the gallbladder. The patient initially elecied treatmant consisling of exage- nously supplied chenadeoxycholic acid, but eventu- ally underwent surgery for the removal of the Correct answer = D. Chenodeoxycholic acid is a hile acid Used in the treatment of gallstones. itis an emphipathic molecule that can act like an emulsifying agent and help solubilize choles- terol, The compound will not affect the entero- hepatic circulation, interlara with cholesterol ‘syntheeis, inctaase bile acid production, or stim- Ulgte VLDL production. gallbladder, and had a full recovery. The rationale for the initial treatment of this patient with chenodeoxy- cholic acid is that this compound: A. interferes with the enterohepatic circulation. inhibits cholesterol synthesis. B. C. increases de novo bile acid production. D. increases cholesterol solubility in bile.244 18, Cholesterol and Steroid Metabolism For Questions 18.2 and 18.3: A young girl with a history of severe abdominal pain was taken to her local hospital at 5 a.m. in severe distress. Blood was drawn, and the plasma appeared milky, with the triacyl- glycerol level in excess of 2,000 mg/dl (normal = 4-150 mg/dl). The patient was placed on a diet severely limited in fat, but supplemented with medium-chain fatty acids. 18.2 Which of the following lipoprotein particles are most likely responsible for the appearance of the patient's plasma? A. Chylomicrons. B. Very-low-density lipoproteins. C. Intermediate-density lipoproteins. D. Low-density-lipoproteins. E. High-density-lipoproteins. 18.3 Medium-chain fatty acids are given because they: A. are more calorically dense than long-chain fatty acids. B. enter directly into the portal blood, and can be metabolized by the liver. . are activators of lipoprotein lipase. . are more efficiently packed into serum lipoproteins. O00E. can be converted into a variety of gluconeogenic precursors. E. stimulates VLDL production by the liver. 18.4 Complete the following table for an individual with classic 21-c-hydroxylase deficiency relative to a nor- mal individual. Aldosterone Cortisol Androstenedione ACTH Blood glucose Blood pressure How might the results be changed if the individual was deficient in 17-o-hydroxylase, rather than 21-c- hydroxylase?Correct answer = A. The milky appearance of her blood was a result of triacylglycerol-rich chy- lomicrons. Because 5 a.m. is presumably sev- eral hours after her evening meal, she must have difficulty clearing these lipoprotein parti- cles. IDL, LDL, or HDL contain primarily choles- teryl esters and, if one or more of these particles was elevated, it would cause hypercholes- terolemia. VLDL do not cause the described “milky appearance” in plasma. Correct answer = B. Medium-chain fatty acids are not packaged in chylomicrons, but rather are carried by albumin to the liver where they can be metabolized. They have the same caloric den- sity as long-chain fatty acids, and are generally much more ketogenic than glycogenic. Lipoprotein lipase does not play a role in their metabolism.Measured Increased] Decreased Aldosterone Cortisol Androstenedione ACTH Blood glucose Blood pressure21-o-Hydroxylase deficiency causes mineralo- corticoids (aldosterone) and glucocorticoids (cortisol) to be virtually absent. Because aldos- terone increases blood pressure, and cortisol increases blood glucose, their deficiencies result in a decrease in blood pressure and blood glu- cose, respectively. Cortisol normally feeds back to inhibit ACTH release by the pituitary, and so its absence results in an elevation in ACTH. The loss of 21-a-hydroxylase pushes progesterone and pregnenolone to androgen synthesis, thus, androstenedione levels rise. With 17-o-hydroxy- lase deficiency, sex hormone synthesis would be inhibited. Mineralocorticoid production would be increased, leading to hypertension.260 19. Amino Acids: Disposal of Nitrogen Study Questions: Choose the ONE best answer. 19.1 In the transamination reaction shown below, which of the following are the products, X and ¥? Oxaloacetate >< X Glutamate Y A. Alanine, o-ketoglutarate. B. Glutamate, o-ketoglutarate. C. Asparate, o-ketoglutarate. D. Pyruvate, aspartate. E. Pyruvate, alanine. 19.2 Which one of the following statements about the urea cycle is correct? A. The two nitrogen atoms that are incorporated into urea enter the cycle as ammonia and alanine.B. Urea is produced directly by the hydrolysis of ornithine. C. ATP is required for the reaction in which argini- nosuccinate is cleaved to form arginine. D. Urinary urea is increased by a diet rich in protein. E. The urea cycle occurs exclusively in the cytosol. For Questions 19.3 and 19.4: 19.3 19.4 A female neonate did well until approximately 24 hours of age when she became lethargic. A sepsis workup proved negative. At 56 hours, she started showing focal seizure activity. The plasma ammonia level was found to be 1,100 mol/L (normal 5-35 umol/L), Quantitative plasma amino acid levels revealed a marked elevation of argininosuccinate. Which one of the following enzymic activities is most likely to be deficient in this patient? A. Arginase. B. Argininosuccinate lyase. C. Argininosuccinate synthase. D. Carbamoyl phosphate synthetase |. E. Ornithine transcarbamoylase. Which one of the following would also be elevated in the blood of this patient?A. Asparagine. B. Glutamine. C. Lysine. D. Urea. E. Uric acid. Correct answer = C. Transaminase reactions always have an amino acid and an a-keto acid as substrates. The products of the reaction are also an amino acid (corresponding to the a-keto substrate) and an o-keto acid (corresponding to the amino acid substrate). Three amino acid a- keto acid pairs commonly encountered in meta- bolism are: alanine/pyruvate aspartate/oxaloacetate glutamate/o-ketoglutarate In this question, glutamate is deaminated to form a-ketoglularate, and oxaloacetate is ami- nated to form aspartate. ~Correct answer = D. The amino nitrogen of dietary protein is excreted as urea. The two nitro- gens enter the urea cycle as ammonia and aspartate. Urea is produced by the hydrolysis of arginine. The cleavage of argininosuccinate does not require ATP. The urea cycle occurs partly in the mitochondria. Nee Correct answer = B. Genetic deficiencies of each of the five enzymes of the urea cycle, as well as deficiencies in N-acetyglutamate synthase, have been described. The accumulation of argini- nosuccinate in the plasma of this patient means that the enzymes required for its synthesis are functional, but the enzyme (argininosuccinate lyase or argininosuccinase) required for its cleav- age to arginine plus fumarate is not. NSS SSS SSCS SO OSS SOOO SSOSU SESS OIRSE SREfeo oe re Correct answer = B. With the exception of arginase, deficiencies of the enzymes of the urea cycle result in the failure to synthesize urea and lead to hyperammonemia in the first few weeks after birth. Glutamine will also be elevated because it acts as a nontoxic storage and trans- port form of ammonia. Thus, elevated glutamine always accompanies hyperammonemia. Asparagine does not serve this sequestering role. Urea would be decreased due to impaired activity of the urea cycle. Lysine and uric acid would not be elevated. Treatment of this patient includes limiting protein in the diet and administering com- pounds that bind covalently to amino acids, pro- ducing nitrogen-containing molecules that are excreted in the urine. For example, phenylbu- tyrate given orally is converted to phenylacetate. This compound condenses with glutamine to form phenylacetylglutamine, which is excreted.276 20. Amino Acid Degradation and Synthesis Study Questions: Choose the ONE correct answer. 20.1 Which one of the following statements concerning amino acids is correct? A. Anincrease in gluconeogenesis from amino acids results in a decrease in urea formation. B. All essential amino acids are glycogenic. C. Omithine and citrulline are found in tissue pro- teins. D. Cysteine is an essential amino acid in individuals consuming a diet severely limited in methionine. E. In the presence of adequate dietary sources of tyrosine, phenylalanine is not an essential amino acid. 20.2 Which one of the following statements concerning a 1-week-old male infant with undetected classic phenylketonuria is correct? A. Tyrosine is a nonessential amino acid for theinfant. . High levels of phenylpyruvate appear in his urine. . Therapy must begin within the first year of life. . A diet devoid of phenylalanine should be initiated immediately. E. When the infant reaches adulthood, it is recom- mended that diet therapy be discontinued. 0oOwn 20.3 A 4-year-old boy of a first-degree consanguineous couple was noted by the parents to have darkening of the urine to an almost black color when it was left standing. He has a normal sibling, and there are no other medical problems. Growth and development to date are normal. Which of the following is most likely to be elevated in this patient? A. Methylmalonate. B. Homogentisate. C. Phenylpyruvate. D. a-Ketoisovalerate. E. Homocysteine. 20.4 What enzyme is deficient in: homocystinuria, methyl- malonic acidemia, MSUD, oculocutaneous albinism, PKU?Correct answer = D. Methionine is the precursor of cysteine. An increase in the availability of glu- coneogenic amino acids from the catabolism of body protein is associated with increased ammonia and results in increased urea produc- tion. The essential amino acids leucine and lysine are ketogenic. Ornithine and citrulline are amino acids that are intermediates in the urea cycle, but are not found in tissue proteins because there are no codons for them. Phenylalanine is essential regardless of the level of tyrosine.Correct answer = B. Phenyllactate, phenyl- acetate, and phenylpyruvate, which are not nor- mally produced in significant amounts in the presence of functional phenylalanine hydroxy- lase, are elevated in PKU, and appear in the urine. In patients with PKU, tyrosine cannot be synthesized from phenylalanine and, hence, becomes essential and must be supplied in the diet. Treatment must begin during the first 7-10 days of life to prevent mental retardation. Discontinuance of the phenylalanine-restricted diet before 8 years of age is associated with poor performance on IQ tests. Adult PKU patients show deterioration of attention and speed of mental processing after discontinuation of the diet. Elevated levels of phenylalanine are teratogenic. Lifelong restriction of dietary pheny- lalanine is, therefore, recommended.Correct answer = B. Alkaptonuria is a rare metabolic disease involving a deficiency in homogentisic acid oxidase, and the subsequent accumulation of homogentisic acid in the urine, which turns dark upon standing. The elevation of methylmalonate (due to methylmalonyl CoA mutase deficiency), phenylpyruvate (due to phenylalanine hydroxylase deficiency), a- ketoisovalerate (due to branched-chain a-keto acid dehydrogenase deficiency), and homocys- teine (due to cystathionine B-synthase defi- ciency) are inconsistent with a healthy child with darkening of the urine. Answer: Cystathionine B-synthase, methyl- malonyl CoA mutase, branched-chain o-keto acid dehydrogenase, tyrosinase, phenylanine hydroxylase. [Note:Deficiency of dihydropteri- dine reductase or any of the enzymes needed for BH4 synthesis can also result in hyperpheny- lalaninemia.]290 21. Conversion of Amino Acids to Specialized Products Study Questions Choose the ONE best answer. 21.1 §-Aminolevulinic acid synthase activity: A. in liver is frequently decreased in individuals treated with drugs, such as the barbiturate pheno- barbital. B. catalyzes a rate-limiting reaction in porphyrin biosynthesis. C. requires the coenzyme biotin. D. is strongly inhibited by heavy metal ions such as lead. E. occurs in the cytosol. 21.2 The catabolism of hemoglobin: A. occurs in red blood cells. B. involves the oxidative cleavage of the porphyrin ring. C. results in the liberation of carbon dioxide. D. results in the formation of protoporphyrinogen. E. is the sole source of bilirubin. 21.3 A 50-year-old man presented with painful blisters on the backs of his hands. He was a golf instructor, andindicated that the blisters had erupted shortly after the golfing season began. He did not have recent exposure to poison ivy or sumac, new soaps or detergents, or new medications. He denied having previous episodes of blistering. He had partial com- plex seizure disorder that had begun about three years earlier after a head injury. The patient had been taking phenytoin—his only medication—since the onset of the seizure disorder. He admitted to an average weekly ethanol intake of about eighteen 12- oz cans of beer. The patient’s urine was reddish orange. Cultures obtained from skin lesions failed to grow organisms. A 24-hour urine collection showed elevated uroporphyrin (1,000 mg; normal, <27mg). The most likely presumptive diagnosis is: A. porphyria cutanea tarda. B. acute intermittent porphyria. C. hereditary coproporphyria. D. congenital erythropoietic porphyria. E. erythropoietic protoporphyria. 21.4 A 10-year-old boy is referred to a specialist because of skin that blisters easily, urine that darkens on standing, and stained teeth. Lab stucies are remark- able for high levels of uroporphyrin | and copropor- phyrin | in plasma, with uroporphyrin | being present in the urine. The most likely biochemical pathology in this case is:A. deficiency of ALA synthase. B. deficiency of bilirubin glucuronyltransferase. C. deficiency of uroporphyrinogen Ill synthase. D. down-regulation of tyrosinase. E. inhibition of ALA dehydratase by lead. Correct answer = B. The activity of 5-aminolev- ulinic acid synthase controls the rate of por- phyrin synthesis. The hepatic form of the enzyme is increased in patients treated with cer- tain drugs, and requires pyridoxal phosphate as a coenzyme. Another enzyme in the pathway (6- aminolevulinic acid dehydrase) is extremely sen- sitive to the presence of heavy metals.Correct answer = B. The cyclic heme molecule is oxidatively cleaved to form biliverdin. The catabolism occurs in the cells of the reticulo- endothelial system, particularly the spleen, and results in the liberation of carbon monoxide. Protoporphyrinogen is an intermediate in the synthesis, not degradation, of heme. Cytochromes and other non-hemoglobin heme- proteins are also precursors of bilirubin. Correct answer = A. The disease is associated with a deficiency in uroporphyrinogen decar- boxylase, but clinical expression of the enzyme deficiency is influenced by hepatic injury causedby iron overload, chronic ethanol consumption, and the presence of hepatitis B or C and HIV infections. Exposure to sunlight can also be a precipitating factor. Clinical onset is typically during the fourth or fifth decade of life. Porphyrin accumulation leads to cutaneous symptoms and urine that is red to brown. Treatment of the patient's seizure disorder with phenytoin caused increased synthesis of ALA synthase, and, therefore, of uroporphyrinogen, the substrate of the deficient enzyme. The laboratory and clinical findings are inconsistent with other porphyrias.Correct answer = C. A deficiency of uropor- phyrinogen III synthase results in accumulation of hydroxymethylbilane and the spontaneous conversion of this substrate to porphyrins of the Type | series. A deficiency of ALA synthase or inhibition of ALA dehycratase by lead would not allow the synthesis of porphobilinogen, the first pyrrole product in the heme biosynthetic path- way, and thus would not result in uro- or copro- porphyrin synthesis. Deficiency of the glucuronyltransferase would not present with the systems described, and lab studies would be remarkable for an elevation of unconjugated bilirubin. Down-regulation of tyrosinase would result in decreased pigmentation.306 22. Nucleotide Metabolism Study Questions Choose the ONE correct answer. 22.1 A 42-year-old male patient undergoing radiation ther- apy for prostate cancer develops severe pain in the metatarsal phalangeal joint of his right big toe. Monosodium urate crystals are detected by polarized light microscopy in fluid obtained from this joint by arthrocentesis. Uric acid crystals are present in his urine, This patient’s pain is directly caused by the overproduction of the end product of which of the fal- lowing metabolic pathways? . De novo pyrimidine biosynthesis. . Pyrimidine degradation. . De novo purine biosynthesis. . Purine salvage. . Purine degradation. moow >, 22.2 A 1-year-old female patient is lethargic, weak, and anemic. Her height and weight are both low for her age. Her urine contains an elevated level of oroticacid. The administration of which of the following compounds is most likely to alleviate her symptoms? A. Adenine. B. Guanine. C. Hypoxanthine. D. Thymidine. E. Uridine. 22.3 The rate of DNA synthesis in a culture of cells could be most accurately determined by measuring the incorporation of which of the following radiolabeled compounds? A. Adenine. B. Guanine. C. Phosphate. D. Thymidine. 22.4 Which one of the following enzymes of nucleotide metabolism is correctly paired with its pharmacologi- cal inhibitor? A. Dihydrofolate reductase—methotrexate B. IMP dehydrogenase—hydroxyurea CG. Ribonucleotide reductase—5-fluorouracil D. Thymidylate synthase—allopurinol E. Xanthine oxidase—probenecid22.5 What laboratory test would help in distinguishing an orotic aciduria caused by ornithine transcarbamylase deficiency from that caused by UMP synthase defi- ciency? Correct answer = E. The patient's pain is caused by gout, resulting from an inflammatory response to the crystallization of excess uric acid in his joints. Radiation therapy caused cell death, with degradation of nucleic acids and their constituent purines. Uric acid, the end product of purine degradation, is a relatively insoluble compound that can cause gout and kidney stones. Pyrimidine metabolism is not associated with uric acid production. Overproduction of purines can indirectly result in hyperuricemia. Purine salvage decreases uric acid production.Le Correct answer = E. The elevated excretion of orotic acid indicates that the patient has orotic aciduria, a rare genetic disorder affecting the de novo pyrimidine biosynthetic pathway. Deficiencies in the enzyme activities OMP decarboxylase and/or orotate phosphoribosyl- transferase (both of which are domains of the enzyme UMP synthase) leave the patient unable to synthesize pyrimidines. Uridine, a pyrimidine nucleoside, is useful in treating this disorder because it bypasses the missing enzymes and can be salvaged to UMP, which can be con- verted to all the other pyrimidines. Although thymidine is a pyrimidine nucleoside, it cannot be converted to other pyrimidines. Hypo- xanthine, guanine, and adenine are all purine bases that have no value in helping to replace the missing pyrimidines.Ge eee ee ee ee ED) Correct answer = D. Because thymidine is essen- tially found only in DNA, its incorporation would most accurately reflect the rate of DNA synthesis. Uridine is found only in RNA and could be used to measure the rate of RNA synthesis. Phosphate, adenine, and guanine are present in both DNA and RNA, and could not be used to specifically measure synthesis of either one. Cn ee rr Correct answer = A. Methotrexate interferes with folate metabolism by acting as a competitive inhibitor of the enzyme dihydrofolate reductase. This starves cells for tetrahydrofolate, and makes them unable to synthesize purines and dTMP. IMP dehydrogenase is inhibited by mycophenolic acid. Ribonucleotide reductase is inhibited by hydroxyurea. Thymidylate synthase is inhibited by 5-fluorouracil. Xanthine oxidase is inhibited by allopurinol; probenecid increases renal excretion of urate, but does not inhibit its production. NEBlood ammonia level would expected to be ele- vated in ornithine transcarbamylase deficiency but not in UMP synthase deficiency.320 23. Metabolic Effects of Insulin and Glucagon Study Questions Choose the ONE correct answer. 23.1 In which one of the following tissues is glucose trans- port into the cell insulin-sensitive? A. Brain. B. Lens of the eye. C. Red blood cells. D. Adipose tissue. E. Liver. 23.2 Which one of the following is characteristic of low insulin levels? A. Increased glycogen synthesis. B. Decreased gluconeogenesis from lactate. C. Decreased glycogenolysis. D. Increased formation of 3-hydroxybutyrate. E. Decreased action of hormone-sensitive lipase.23.3 Which one of the following statements about glucagon is correct? A. High levels of blood glucose increase the release of glucagon from the « cells of the pancreas. B. Glucagon levels decrease following ingestion of a protein-rich meal. C. Glucagon increases the intracellular levels of cAMP in liver cells, causing an increase in glycogenolysis. D. Glucagon is the only hormone important in combat- ing hypoglycemia. E. Glucagon depresses the formation of ketone bod- ies by the liver. 23.4 A 39-year-old woman is brought to the emergency room complaining of dizziness. She recalls getting up early that moming to do as much shopping as possible and had skipped breakfast. She drank a cup of coffee for lunch and had nothing to eat during the day. She met with friends at 8 p.m. and had a drink at the bar. She soon became weak and dizzy and was trans- ported to the hospital. Following examination, the patient was given orange juice and immediately felt better. Which one of the following best completes this sentence? “The patient has”: A. blood glucose greater than 70 mg/dl. B. elevated insulin.C. elevated glucagon. D. elevated liver glycogen. E. presence of an insulinoma. Correct answer = D. The major tissues in which glucose transport requires insulin are muscle and adipose tissue. The metabolism of the liver responds to insulin, but hepatic glucose transport is determined by blood glucose concentration and does not require insulin. Brain, RBC, and lens of the eye have insulin-insensitive uptake of glucose. Correct answer = D. 3-Hydroxybutyrate (or f- hydroxybutyrate)—a ketone body—synthesis is enhanced in the liver by low insulin levels, which favor activation of hormone-sensitive lipase and release of fatty acids from adipose tissue. Glycogen synthesis is decreased, whereas gluco- neogenesis and glycogenolysis are increased.Correct answer = C. The cAMP cascade initiated by glucagon causes the liver to degrade glyco- gen, releasing glucose to the blood. High levels of blood glucose decrease the release of glucagon from the o cells of the pancreas. Glucagon levels increase following ingestion of a protein-rich meal. In addition to glucagon, epi- nephrine and cortisol are also important in increasing glucose production in hypoglycemia. Glucagon increases the formation of ketone bodies by the liver. Correct answer = C. The patient's glucagon level will be elevated in response to the hypoglycemia. She is most likely experiencing alcohol-induced fasting hypoglycemia. Blood glucose is expected to be 40 mg/dl or less, insulin secretion depres- sed because of the low blood glucose, and liver glycogen levels low because of the fast. An insulinoma, an insulin-producing tumor of the pancreas, is unlikely.336 24. The Feed/Fast Cycle Study Questions Choose the ONE correct answer. 24.1 Which one of the following is elevated in plasma during the absorptive (fed) period as compared with the postabsorptive (fasted) state? A. Glucagon. B. Acetoacetate. C. Chylomicrons. D. Free fatty acids. E. Lactate. 24,2 Which one of the following statements concerning the fed state is correct? A. Most enzymes that are regulated by covalent modification are in the phosphorylated state. B. Hepatic fructose 2,6-bisphosphate is elevated. C. The oxidation of acetyl CoA is increased. D. Insulin stimulates the transport of glucose into hepatocytes.24.3 24.4 E. The synthesis of glucokinase is repressed. Increased formation of ketone bodies during fasting is a result of: A. decreased levels of circulating glucagon. B. decreased formation of acetyl CoA in the liver. C. increased levels of free fatty acids in blood. D. inhibition of B-oxidation of fatty acids in the liver. E. decreased activity of hormone-sensitive lipase in adipose tissue. Which one of the following is the most important source of blood glucose during the last hours of a 48-hour fast? A. Muscle glycogen. B. Acetoacetate. C. Liver glycogen. D. Amino acids. E. Lactate.Correct answer = C. TAG-rich chylomicrons are synthesized in (and released from) the intestine following ingestion of a meal. Glucagon is depressed in the absorptive period. Acetoacetate, free fatty acids, and lactate are not elevated. Correct answer = B. The increased insulin and decreased glucagon levels characteristic of the fed state promote the synthesis of fructose 2,6- bisphosphate. Most covalently modified enzymes are in the dephosphorylated state and are active. Acetyl CoA is not elevated in the fed state. The transport of glucose in the liver is not insulin sensitive. Synthesis of glucokinase is enhanced in the fed state. (en) Correct answer = C. Free fatty acids bound to albumin are increased as a result of an increased activity of hormone-sensitive lipase in adipose tissue. Hepatic ketogenesis is stimu-lated by elevated levels of glucagon. The forma- tion of acetyl CoA is increased. B-Oxidation of fatty acids in liver provides the acetyl CoA for ketogenesis. Correct answer = D. The carbon skeletons of glucogenic amino acids are used by the liver for gluconeogenesis. Liver glycogen is nearly depleted by 12 hours after a meal, and muscle glycogen cannot give rise to free glucose because muscle lacks glucose 6-phosphatase. Acetoacetate is metabolized to acetyl CoA, which is not glucogenic. Lactate can arise from anaero- bic glycolysis in muscle and red blood cells, but is less important than amino acids as a source of glucose. ee348 25. Diabetes Mellitus Study Questions Choose the ONE correct answer. 25.1 Relative or absolute lack of insulin in humans would result in which one of the following reactions in the liver? A. Increased glycogen synthesis. B. Decreased gluconeogenesis from lactate. C. Decreased glycogenolysis. D. Increased formation of 3-hydroxybutyrate. E. Decreased action of hormone-sensitive lipase. 25.2 Which one of the following is most often found in untreated patients with type 1 and type 2 diabetes? A. Hyperglycemia. B. Extremely low levels of insulin synthesis and secretion. C. Synthesis of an insulin with an abnormal amino acid sequence. D. A simple pattern of genetic inheritance. E. Ketoacidosis.25.3 An obese individual with type 2 diabetes: 25.4 A. B. Cc. usually shows a sudden onset of symptoms. usually has a lower plasma level of insulin than a normal individual. usually shows significant improvement in glucose tolerance if body weight is reduced to normal. . usually benefits from receiving insulin about 6 hours after a meal. . usually has lower plasma levels of glucagon than a normal individual. An individual with insulin resistance and normal B- cell function: mooopr . usually shows elevated fasting glucose levels. . usually shows elevated fasting insulin levels. . will eventually become diabetic. . is rarely obese. . is treated by injection of insulin. 25.5 Explain why drugs that inhibit the a-glucosidase activity of the intestinal saccharidases aid in glycemic control in patients with diabetes.Correct answer = D. Low insulin levels favor the liver producing ketone bodies, using acetyl coenzyme A it generated by B-oxidation of fatty acids provided by the adipose. Low insulin also causes activation of hormone-sensitive lipase, decreased glycogen synthesis, and increased gluconeogenesis and glycogenolysis. Correct answer = A. Elevated blood glucose occurs in type 1 diabetes as a result of a lack of insulin. In type 2 diabetes, hyperglycemia is due to a defect in B-cell function and insulin resistance. The amino acid sequence of insulin is not changed in diabetes. Both forms of the disease show complex genetics. Ketoacidosis is more common in type 1 disease.Correct answer = C. Eighty percent of type 2 diabetics are obese, and almost all show some improvement in blood glucose with weight reduction. Symptoms usually develop gradually. These patients have elevated insulin levels, and usually do not require insulin (certainly not 6 hours after a meal). Glucagon levels are typi- cally normal. Correct answer = B. Insulin resistance is the decreased ability of target tissues, such as liver, adipose, and muscle, to respond properly to nor- mal circulating concentrations of insulin. Obesity is the most common cause of insulin resistance. Most of the people with obesity and insulin resis- tance do not become diabetic. In the absence of a defect in B-cell function, nondiabetic, obese individuals can compensate for insulin resis- tance with elevated levels of insulin. The ele- vated insulin levels normalize fasting blood glucose levels. Insulin resistance without overt diabetes requires no pharmacologic treatment.o-Glucosidase inhibitors prevent glucose produc- tion from those products of carbohydrate diges- tion in which glucose is attached through an o-glycosidic linkage, thus reducing the postpran- dial rise in blood glucose. Note that the digestion of lactose is unaffected because it has a B-link- age.356 26. Obesity Study Question Choose the ONE correct answer. 26.1 A 40-year-old woman, 5 feet, 1 inch (155 cm) tall and weighing 188 pounds (85.5 kg), seeks your advice on how to lose weight. Her waist measured 41 inches and her hips 39 inches. A physical examination and blood laboratory data were all within the normal range. Her only child, who is 14 years old, her sister, and both of her parents are overweight. The patient recalls being obese throughout her childhood and adolescence. Over the past 15 years she had been on seven different diets for periods of 2 weeks to 3 months, losing from 5-25 pounds. On discontinuation of each diet, she regained weight, returning to 185-190 pounds. Which one of the following best describes this patient? A. She is classified as overweight. B. She shows an “apple” pattern of fat distribution. C. She has approximately the same number of fat cells as a normal-weight individual, but each adipocyte is larger.D. She would be expected to show lower than nor- mal levels of circulating leptin. E. She would be expected to show lower than nor- mal levels of circulating triacylglycerols. 26.2 Calculate the BMI for the patient presented in ques- tion 26.1Gorrect answer = B. Her waist to hip ratio is 41/39 = 1.05. Apple shape is defined as a waist to hip ratio of more than 0.8 for women, and more than 1.0 for men. She has, therefore, an apple pattern of fat distribution, more commonly seen in males. Compared with other women of the same body weight who have a gynoid fat pattern, the presence of increased visceral or intra-abdominal adipose tissue places her at greater risk for diabetes, hypertension, dyslipi- demia, and coronary heart disease. The calcu- lated BMI indicates that the patient is classified as obese. Individuals with marked obesity and a history dating to early childhood have an adi- pose depot made up of too many adipocytes, each fully loaded with triacylglycerols. Plasma leptin in obese humans is usually normal for their fat mass, suggesting that resistance to lep- tin, rather than its deficiency, occurs in human obesity. The elevated circulating fatty acids characteristic of obesity are carried to the liver and converted to triacylglycerol and cholesterol. Excess triacylglycerol and cholesterol are released as VLDL, resulting in elevated serum triacylglycerols.BMI = weight (kg)/height (m2) = 85.5/(1.55)? = 35.6 kg/m?.370 27. Nutrition Study Questions Choose the ONE correct answer. 27.1 Which one of the following statements concerning ‘dietary lipid Is correct? A. Corn oll and soybean oil are examples of fats rich in saturated fatty acids. B. Triacylglycerols obtained from plants generally contain less unsaturated fatty acids than those from animals. C. Olive oll Is rich In saturated fats. Correct answer = D. Trans fatty acids raise plasma cholesterol levels, Corn cil and soybean all are examples of fats rich In polyunsaturated {ally acids, Trlacylglyeerols. oblalned from planis generally contain more unsaturated fatty acids than those from animals. Olive oil, the staple of the Mediterranean diet, is rich in moncunsetu- tated fats. Coconut and palm olis are unusual plant olis in that they are nici in saturated fats, D. Fatty acids containing double bonds in the trans configuration, unlike the naturally occurring cis isomers, raise plasma cholesterol levels. E. Coconut and palm oils are rich in polyunsaturated fats.372 27. Nutrition 27.2 27.3 Given the information that a 70-kg man is consuming a daily average of 275 g of carbohydrate, 75 g of pro- tein, and 65 g of lipid, one can draw which of the fol- lowing conclusions? A. Total energy intake per day is approximately 3,000 keal. B. About 20% of the calories are derived from lipids. C. The diet does not contain a sufficient amount of dietary fiber. D. The proportions of carbohydrate, protein, and lipid in the diet conform to the recommendations of academic groups and government agencies. E. The individual is in nitrogen balance, A sedentary 50-year-old man, weighing 80 kg (176 pounds), requests a physical examination. He denies any health problems. Routine blood analysis is unre- markable except for plasma cholesterol of 280 mg/dl. The man refuses drug therapy for his hypercholes- terolemia. Analysis of a 1-day dietary recall showed the following: Kilocalories 3,475 kcal Cholesterol 822 mg Protein 102 g Saturated fat 69g Carbohydrate 383g Total Fat 1659Fiber crude 6g Changes in which one of the following dietary compo- nents would have the greatest effect in lowering plasma cholesterol? A. Cholesterol. B. Saturated fat. C. Polyunsaturated fat. D. Monounsaturated fat. E. Carbohydrate. 27.4 Which of the findings in the child shown at right would support a diagnosis of kwashiorkor? A. Shows increased serum alburnin. B. Shows a good appetite, C. Appears plump due to increased adipose tissue. D. Displays abdominal and peripheral edema. E, Has markedly decreased weight for height,Correct answer = D. The total energy intake is (275 g carbohydrate x 4 kcal/g) + (75 g protein x 4 kcal/g) + (65 g lipid x 9 keal/g) = 1100 + 300 + 585 = 1,985 total kcal/day. The percentage calo- ries from carbohydrate is 1,100/1,985 = 55; per- centage calories from protein is 300/1,985 = 15; and percentage calories derived from lipid is 585/1,985 = 30. These are very close to current recommendations, The amount of fiber or nitro- gen balance cannot be deduced from the data presented. If the protein is of low biologic value, a negative nitrogen balance is possible.Correct answer = B. The intake of saturated fat most strongly influences plasma cholesterol in this diet. The patient is consuming a high-calorie, high-fat diet with 40% of the fat as saturated fat. The most important dietary recommendations are: lower total caloric intake, substitute monoun- saturated and polyunsaturated fats for saturated fats, and increase dietary fiber. A decrease in dietary cholesterol would be helpful, but is not a primary objective.The correct answer = D. Kwash- iorkor is caused by inadequate pro- tein intake in the presence of fair to good energy (calorie) intake. Typical findings in a patient with Kwashiorkor include abdominal and peripheral edema (note the swollen belly and legs of the child) caused largely by a decreased serum albu- min concentration. Anorexia is almost always present. Weight for height can be normal. Treatment includes a diet adequate in calories and protein.394 28. Vitamins Study Questions Choose the ONE correct answer. 28.1 Which one of the following statements concerning vitamin Bia is correct? 28.2 mo oof . The cofactor form is vitamin B,> itself. . Itis involved in the transfer of amino groups. . It requires a specific glycoprotein for its absorp- tion. . Itis present in plant products. . Its deficiency is most often caused by a lack of the vitamin in the diet. Retinol: A. B. Cc. can be enzymically formed from retinoic acid, in its ester form is transported from the intestine to the liver in chylomicrons. is the light-absorbing portion of rhodopsin. is phosphorylated and dephosphorylated during the visual cycle. . mediates most of the actions of the retinoids.28.3 Which one of the following statements concerning 28.4 28.5 vitamin D is correct? A. Chronic renal failure requires the oral administra- tion of 1,25-dihydroxycholecalciferol. B. It is required in the diet of individuals exposed to sunlight. C.25-Hydroxycholecalciferol is the active form of the vitamin. D.Vitamin D opposes the effect of parathyroid hor- mone. E.A deficiency in vitamin D results in an increased secretion of calcitonin. Vitamin K: A. plays an essential role in preventing thrombosis. B. increases the coagulation time in newborn infants with hemorrhagic disease. C. is present in high concentration in cow or breast milk. D. is synthesized by intestinal bacteria. E. is a water-soluble vitamin. Deficiency of which vitamin results in: beri-beri; scurvy; pellagra; night blindness; rickets or osteoma- lacia; pernicious anemia; megaloblastic anemia;XN bleeding? Correct answer = C. Vitamin Bi2 requires intrinsic factor for its absorption. A deficiency of vitamin Bis is most often caused by a lack of intrinsic fac- tor. However, high does of the vitamin, given orally, are sufficiently absorbed to serve as treat- ment for pernicious anemia. The cofactor forms are methylcobalamin and deoxyadenosylcobal- amin. Vitamin Bg, not vitamin By, is involved in the transfer of amino groups. By2 is found in food derived from animal sources. Correct answer = B. Retinyl esters are incorpo- rated into chylomicrons. Retinoic acid cannot be reduced to retinol. Retinal, the aldehyde form of retinol, is the chromophore for rhodopsin. Retinal is photoisomerized during the visual cycle. Retinoic acid, not retinol, is the most important retinoid.Correct answer = A. Renal failure results in the decreased ability to form the active form of the vitamin, which must be supplied. The vitamin is not required in individuals exposed to sunlight. 1,25-Dihydroxycholecalciferol is the active form of the vitamin. Vitamin D and parathyroid hormone both increase serum calcium. A deficiency of vita- min D decreases the secretion of calcitonin. \ Correct answer = D. Vitamin K is essential for clot formation, decreases coagulation time, and is present in low concentrations in milk. It is one of four fat-soluble vitamins. / Thiamin; vitamin ©; niacin; vitamin A, vitamin D; vitamin B12 (due primarily to a deficiency of intrinsic factor), both B12 and folate; vitamin K. *416 29. DNA Structure, Replication, and Repair Study Questions Choose the ONE correct answer. 29.1 A 10-year-old girl is brought to the dermatologist by her parents. She has many freckles on her face, neck, arms, and hands, and the parents report that she is unusually sensitive to sunlight. Two basal cell carcinomas are identified on her face. Which of the following processes is most likely to be defective in this patient? A. Repair of double-strand breaks. B. Removal of mismatched bases from the 3'-end of Okazaki fragments. C. Removal of pyrimidine dimers from DNA. D. Removal of uracil from DNA. 29.2 Telomeres are complexes of DNA and protein that protect the ends of linear chromosomes. In most nor- mal human somatic cells, telomeres shorten with each division. In stem cells and in cancer cells, how- ever, telomeric length is maintained. In the synthesisof telomeres: A. telomerase, a ribonucleoprotein, provides both the RNA and the polymerase needed for synthesis. B. the RNA of telomerase serves as a primer. C. the polymerase of telomerase is a DNA-directed DNA polymerase. D. the shorter, 3'—5' strand gets extended. E. the direction of synthesis is 3'35'. 29.3 While studying the structure of a small gene that was recently sequenced during the Human Genome Project, an investigator notices that one strand of the DNA molecule contains 20 As, 25 Gs, 30 Cs, and 22 Ts. How many of each base is found in the complete double-stranded molecule? A. A= 40, G=50, C = 60, T = 44. B. A= 44, G = 60, C = 50, T = 40. C. A=45, G=45, C = 52, T=52. D. A=50, G=47, C= 50, T =47. E. A= 42, G = 55, C= 55, T = 42. 29.4 The extent of DNA synthesis in a cell could most specifically be determined by measuring the incorpo- ration of radiolabeled: A. leucine. B. phosphate. C. riboseD. thymidine E. uracil Correct answer = C. The sensitivity to sunlight, extensive freckling on parts of the body exposed to the sun, and presence of skin can- cer at a young age indicates that the patient most likely suffers from xeroderma pigmento- sum. These patients are deficient in any one of several XP proteins required for nucleotide excision repair of ultraviolet-damaged DNA. Double-strand breaks are repaired by non- homologous end-joining or homologous recom- bination. Uracil is removed from damaged DNA molecules by a specific glycosylase. ee ee en Correct answer = A. Telomerase is a ribonucleo- protein particle required for telomere mainte- nance. Telomerase contains an RNA that serves as the template, not the primer, for the synthesis of telomeric DNA by the reverse transcriptase oftelomerase. As a reverse transcriptase, it syn- thesizes DNA using its RNA template and so is an RNA-directed DNA polymerase. The direc- tion of synthesis, as with all DNA synthesis, is 5'33', and it is the already longer 5'—3' strand that gets extended. Correct answer = E. The two DNA strands are complementary to each other, with A base-paired with T, and G base-paired with C. So, for exam- ple, the 20 As on the first strand would be paired with 20 Ts on the second strand, the 25 Gs on the first strand would be paired with 25 Gs on the second strand, and so forth. When these are all added together, the correct numbers of each base are indicated in choice E. Notice that, in the correct answer, A= T and G=C.Correct answer = D. Thymidine is incorporated only into DNA, and thus would reflect the extent of DNA synthesis. Leucine is an amino acid, nota nucleotide base. Phosphate is not exclusive to DNA. Ribose is found in RNA, whereas deoxyri- bose is found in DNA. Uracil is normally found only in RNA. If present in DNA, likely as a result of deamination of cytosine, it is removed by uracil glycosylase during base excision repair.430 30. RNA Structure, Synthesis, and Repair Study Questions Choose the ONE correct answer. 30.1 A 1-year-old male with chronic anemia is found to have B-thalassemia. Genetic analysis shows that one of his B-globin genes has a mutation that creates a new splice acceptor site 19 nucleotides upstream of the normal splice acceptor site of the first intron. Which of the fol- lowing best describes the new mRNA molecule that can be produced from this mutant gene? A. Exon 1 will be too short. B. Exon 1 will be too long. C. Exon 2 will be too short. D. Exon 2 will be too long. E. Exon 2 will be missing. 30.2 The base sequence of the strand of DNA used as the template for transcription is GATCTAG. What is the base sequence of the RNA product? (All sequences are written 5'- 3' according to standard convention.) A. CTAGATG. B. GTAGATC. C. GAUCUAC.D. CUAGAUG. E. GUAGALC. 30.3 A 4-year-old child who becomes easily tired and has trouble walking is diagnosed with Duchenne muscular dystrophy, an X-linked recessive disorder. Genetic anal- ysis shows that the patient’s gene for the muscle protein dystrophin contains a mutation in its promoter region. Of the choices listed, which would be the most likely effect of this mutation? A. Initiation of dystrophin transcription will be defective. B. Termination of dystrophin transcription will be defec- tive. C. Capping of dystrophin mANA will be defective. D. Splicing of dystrophin mRNA will be defective. E. Tailing of dystrophin mRNA will be defective. 30.4 A mutation to this sequence in eukaryotic mRNA will affect the process by which the 3'-end poly-A tail is added to the mRNA. A. CAAT B. CCA C. GGGGCG D. AAUAAA E. TATAAACorrect answer = D. Because the mutation cre- ates an additional splice acceptor site (the 3'- end) upstream of the normal acceptor site of intron 1, the 19 nucleotides that are usually found at the 3'-end of the excised intron 1 lariat can remain behind as part of exon 2. Exon 2 can, therefore, have these extra 19 nucleotides at its 5'-end. The presence of these extra nucleotides in the coding region of the mutant mRNA molecule will prevent the ribosome from translating the message into a normal P-globin protein molecule. Those mRNA for which the normal splice site is used to remove the first intron will be normal, and their translation will produce normal f-globin protein. Nefo) Correct answer = E. The RNA product has a sequence that is complementary to the template strand and identical to the coding strand of DNA. Uracil (U) is found in RNA in place of the thymine (T) in DNA. Thus, the DNA template 5'- GATCTAC-3' would produce the RNA product 3'-CUAGAUG-5' or, written correctly in the stan- dard direction, 5'-GUAGAUC-3'. \ Correct answer = A. Mutations in the promoter typically prevent formation of the RNA poly- merase || transcription complex, resulting in a decrease in the initiation of mRNA synthesis. A deficiency of dystrophin MRNA will result in a deficiency in the production of the dystrophin protein. Capping, splicing and tailing defects are not a consequence of promoter mutations. They can, however, result in MRNA with decreased stability (capping and tailing defects), or a mRNA in which too many or too few introns have been removed (splicing defects)Correct answer = D. An endonuclease cleaves mRNA just downstream of this polyadenylation signal, creating a new 3'-end to which the poi A polymerase adds the poly-A tail using ATP as the substrate in a template-independent process. CAAT, GGGGCGT, and TATAAA are sequences found in promoters for RNA polymerase Il. CCA is added to the 3'-end of tRNA by nuc/eotidyi transferase.VII. Chapter Summary 447 Study Questions Choose the ONE correct answer. 31.1 A 20-year-old anemic man is found to have an abnor- mal form of B-globin (Hemoglobin Constant Spring) that is 172 amino acids long, rather than the 141 found in the normal protein. Which of the following point mutations is consistent with this abnormality? A. UAA > CAA. B, UAA > UAG. C. CGA > UGA. D. GAU > GAC. E. GCA > GAA. 31.2 A pharmaceutical company is studying a new antibi- otic that inhibits bacterial protein synthesis. When this antibiotic is added to an in vitro protein synthesis system that is translating the mRNA sequence AUGUUUUUUUAG, the only product formed is the dipeptide fMet-Phe. What step in protein synthesis is most likely inhibited by the antibiotic?A. Initiation. B. Binding of charged tRNA to the ribosomal A site. C. Peptidyltransferase activity. D. Ribosomal translocation. E. Termination. 31.3 A tRNA molecule that is supposed to carry cysteine (tRNA *) is mischarged, so that it actually carries alanine (ala-tRNA‘S). Assuming no correction occurs, what will be the fate of this alanine residue during protein synthesis? A. It will be incorporated into a protein in response to an alanine codon. B. It will be incorporated into a protein in response to a cysteine codon. C. It will remain attached to the tRNA, as it cannot be used for protein synthesis. D. It will be incorporated randomly at any codon. E. It will be chemically converted to cysteine by cellu- lar enzymes. 31.4 In a patient with cystic fibrosis caused by the AF508 mutation, the mutant cystic fibrosis transmembrane conductance regulator (CFTR) protein folds incor- rectly. The patient’s cells modify this abnormal pro- tein by attaching ubiquitin molecules to it. What is the fate of this modified CFTR protein?A. It performs its normal function, as the ubiquitin largely corrects for the effect of the mutation. B. Itis secreted from the cell. C. It is placed into storage vesicles. D. It is degraded by the proteasome. E. It is repaired by cellular enzymes. Correct answer = A. Mutating the normal stop codon for B-globin from UAA to CAA causes the ribosome to insert a glutamine at that point. It will continue extending the protein chain until it comes upon the next stop codon further down the message, resulting in an abnormally long protein. A change from UAA to UAG would sim- ply change one termination codon for another and would have no effect on the protein. The replacement of CGA (arginine) with UGA (stop) would cause the protein to be too short. GAU and GAC both encode aspartate and would cause no change in the protein. Changing GCA (alanine) to GAA (glutamate) would not change the size of the protein product.Correct answer = D. Because fMet-Phe is made, the ribosomes must be able to complete initia- tion, bind Phe-tRNA to the A site, and use pep- tidyltransferase activity to form the first peptide bond. Because the ribosome is not able to pro- ceed any further, ribosomal movement (translo- cation) is most likely the inhibited step. The ribosome is, therefore, frozen before it reaches the termination codon of this message. Correct answer = B. Once an amino acid is attached to a tRNA molecule, only the anticodon of that tRNA determines the specificity of incor- poration. The mischarged alanine will, therefore, be incorporated in the protein at a position determined by a cysteine codon.Correct answer = D. Ubiquitination usually marks old, damaged, or misfolded proteins for destruction by the proteasome. There is no known cellular mechanism for repair of dam- aged proteins.448 31. Protein Synthesis 31.5 Many antimicrobials inhibit protein translation. Which of the following antimicrobials is correctly paired with its mechanism of action? A. Tetracyclines inhibit peptidyltransferase. B. Diphtheria toxin binds to the 30S ribosomal sub- unit. C. Puromycin inactivates EF-2. D. Clindamycin binds to the 30S ribosomal subunit. E. Erythromycin binds to the 50S ribosomal subunit. 31.6 Translation of a synthetic polyribonucleotide contain- ing the repeating sequence CAA in a cell-free pro- tein-synthesizing system produces three homopolypeptides: polyglutamine, polyasparagine, and polythreonine. If the codon for glutamine and asparagine are CAA and AAC, respectively, which of the following triplets is the codon for threonine? A. AAC. B. CAA. C. CAC. D. CCA.E. ACA. 31.7 Which of the following is required for both prokaryotic and eukaryotic protein synthesis? A. B. Cc. D. E. Binding of the small ribosomal subunit to the Shine-Dalgarno sequence. fMet~tRNA. Movement of the mRNA out of the nucleus and into the cytoplasm. Recognition of the 5'-cap by initiation factors. Translocation of the peptidyl-tRNA from the A site to the P site. 31.8 Why is the genetic code described as both degener- ate and unambiguous? 31.9 o1-Antitrypsin (A1AT) deficiency can result in emphy- sema, a lung pathology, because the action of elas- tase, a serine protease, is unopposed. Deficiency of A1AT in the lungs is the consequence of its retention in (rather than secretion from) the liver, the site of its synthesis. Proteins such as A1AT that are designed to be secreted are best characterized by which of the following statements? A. B. Their synthesis is initiated on the smooth endo- plasmic reticulum. They contain a mannose 6-phosphate targeting signal.C. They always contain methionine as the N-terminal amino acid. D. They are produced from translation products that have an N-terminal hydrophobic signal sequence. E. They contain no sugars with O-glycosidic linkages because their synthesis does not involve the Golgi apparatus. Correct answer = E. Erythromycin and clin- damycin both bind the 50S ribosomal subunit. Tetracyclines inhibit the 30S ribosomal subunit (chloramphenicol inhibits peptidyl transferase). Diphtheria toxin binds EF-2, inactivating it through ADP ribosylation. Puromycin has struc- ture similar to that of aminoacyl-tRNA. It is incor- porated into the growing chain, and inhibits L further elongation of the peptide chain. 3 (ee) Correct answer = E. The synthetic polynu- cleotide sequence of CAACAACAACAA . . could be read by the in vitro protein synthesizing sys- tem starting at the first C, the first A, or the sec- ond A. In the first case, the first triplet codonwould be CAA, which codes glutamine; in the second case, the first triplet codon would be AAC, which codes for asparagine; in the last case, the first triplet codon would be ACA, which codes for threonine. Ne Si ne a a a en rN Correct answer = E. In both prokaryotes and eukaryotes, continued translation (elongation) requires movement of the peptidyl-tRNA from the A site to the P site to allow the next aminoa- cyl-tRNA to enter the A site. Only prokaryotes have a Shine-Dalgarno sequence and use fMet, whereas only eukaryotes have a nucleus and co- and posttranscriptionally process their mRNA. "SS SS SS SS SSR SE SSS US aE SSMS eee ae ane nO ENE DUTT A given amino acid can be coded for by more than one codon (degenerate code), but a given codon codes for just one particular amino acid (unambiguous code). eeAnswer = D. Synthesis of secreted proteins is begun on free (cytosolic) ribosomes. As the N- terminal hydrophobic signal sequence of the peptide emerges from the ribosome, it is bound by the signal recognition particle, taken to the rough endoplasmic reticulum (REAR), threaded into the lumen, and removed as translation con- tinues. The proteins move through the RER and the Golgi, and undergo processing such as N- glycosylation (RER) and O-glycosylation (Golgi). In the Golgi, they are packaged in secretory vesicles and released from the cell. The smooth endoplasmic reticulum is associated with syn- thesis of lipids, not proteins, and has no ribo- somes attached. Phosphorylation at carbon 6 of terminal mannose residues in glycoproteins tar- gets these proteins (acid hydrolases) to lyso- somes. The N-terminal methionine is removed from most proteins during processing.462 32. Regulation of Gene Expression Study Questions Choose the one correct answer. 32.1 Which of the following mutations is most likely to result in reduced expression of the lac operon? A. i” (no repressor protein made) B. 0° (operator cannot bind repressor protein) C. Cyar (no adenylyl cyclase made) D. Functionally impaired glucose transporterCorrect answer = C. In the absence of glucose, adenylyl cyclase makes cAMP, which forms a complex with the CAP protein. The cAMP—CAP complex binds the CAP site on the DNA, caus- ing RNA polymerase to bind more efficiently to the lac operon promoter, thus increasing expres- sion of the operon. With cya” mutations, adeny- lyl cyclase is not made, and so the operon is unable to be turned on even when glucose is absent and lactose is present. The absence of a repressor protein or decreased ability of the repressor to bind the operator results in constitu- tive (constant) expression of the lac operon.464 32. Regulation of Gene Expression 32.2 Which of the following is best described as being trans-acting? A. CAP site B. Operator C. Promoter D. Repressor 32.3 Which of the following is the basis for the intestine- specific expression of apoprotein B-48? A. DNA rearrangement and loss B. DNA transposition C. RNA alternative splicing D. RNA editing E. RNA interference 32.4 Which of the following is most likely to be true in hemochromatosis, a disease of iron accumulation in the body? A. The mRNA for the transferrin receptor (TfA) is sta- bilized by the binding of IRPs to 3' stem-loop structures known as IREs. B. The mRNA for the TFR is not bound by IRPs, andis rapidly degraded, C. The mRNA for apoferritin is not bound by IRPs at its 5' stem-loop IRE, and is translated. D. The mRNA for apoferritin is bound by IRPs, and is not translated. E. Both B and C 32.5 After several weeks of chemotherapy with methotrex- ate, a cancer patient’s tumor begins to show signs of resistance to treatment. Which of the following mech- anisms is most likely to explain this resistance to methotrexate? A. Overproduction of dihydrofolate reductase. B. Overproduction of xanthine oxidase. C. Deficiency of PRPP synthase. D. Deficiency of thymidine kinase. E. Deficiency of thymidylate synthase. 32.6 The ZYA region of the lac operon will be efficiently transcribed if: A. glucose and lactose are available. CAMP levels are low. B. C. the operator is mutated and can’t be bound by the repressor. D. the repressor is mutated and can’t be bound by the inducer.Correct answer = D. A repressor is a molecule that transits to the DNA, binds, and reduces the expression of that DNA; it is said to be trans- acting. The CAP site, operator, and promoter are part of the DNA itself, and so are said to be cis-acting. Correct answer = D. The production of apoB-48 in the intestine and apoB-100 in liver is the result of RNA editing in the intestine, where a sense codon is changed to a nonsense codon by posttranscriptional deamination of C to U. DNA rearrangement and transposition, as well as RNA interference and alternate splicing, do alter gene expression, but are not the basis of apoB-48 tissue-specific production. Ne Correct answer = E. When iron levels in the body are high, as is seen with hemochromatosis, there is increased synthesis of the iron-storagemolecule, apoferritin, and decreased synthesis of the transferrin receptor (TfR) that mediates iron uptake by cells. These effects are the result of trans-acting iron regulatory proteins (IRPs) bind- ing iron rather than binding cis-acting iron- responsive elements (IREs), resulting in degradation of the mRNA for TfR, and increased translation of the mRNA for apoferritin. No ns ncn coe Neen nS un SLES SLRS) {ey Gorrect answer = A. Methotrexate interferes with folate metabolism by acting as a competi- tive inhibitor of the enzyme dihydrofolate reduc- tase. This starves cells for tetrahydrofolate, and makes them unable to synthesize purines and dTMP. This is especially toxic to rapidly growing cancer cells. Overproduction of dihydrofolate reductase, usually caused by amplification of its gene, can overcome the inhibition of the enzyme at the methotrexate concentrations used for chemotherapy, and can result in resis- tance of the tumor to treatment by this drug.Correct answer = C. The lac operon is nega- tively regulated by the repressor protein binding to the operator region and preventing RNA polymerase from transcribing the Z, Y, and A genes of the operon. If glucose is present, the operon is off as a result of catabolite repres- sion. It is only when glucose is gone, cAMP lev- els are increased, and lactose is available that the operon is maximally induced. If the inducer can’t bind the repressor, the repressor binds the operator and represses transcription.XI. Chapter Summary 487 Study Questions Choose the ONE correct answer. 33.1 Hindlll is a restriction endonuclease commonly used to cut human DNA into pieces before inserting it into a plasmid. Which of the following is most likely to be the recognition sequence for this enzyme? A. AAGGAA. B. AAGAAG. C. AAGTTC. D. AAGCTT. E. AAGAGA.Correct answer = D. The vast majority of restric- tion endonucleases recognize palindromes, and AAGCTT is the only palindrome among the choices. Because the sequence of only one DNA strand is given, one must determine the base sequence of the complementary strand. To be a palindrome, both strands must have the same sequence when read in the 5'>3' direc- tion. Thus, the complement of 5'-AAGCTT-3' is also 5'-AAGCTT-3'.488 33. Biotechnology and Human Disease 83.2 An Ashkenazi Jewish couple brings their 6-month-old son to you for evaluation of listlessness, poor head control, and a fixed gaze. You determine that he has Tay-Sachs disease, an autosomal recessive disor- der. The couple also has a daughter. The diagram below shows this family’s pedigree, along with Southern blots of an RFLP very closely linked to the hexosaminidase A gene, which is defective in Tay- Sachs. Which of the statements below is most accu- rate with respect to the daughter? Father Mother Son Daughter _ — — | 4kb — — — 3kb A. She has a 25% chance of having Tay-Sachs dis-ease. . She has a 50% chance of having Tay-Sachs dis- ease. . She has Tay-Sachs disease. . She is a carrier for Tay-Sachs disease. . She is homozygous normal. moo OD 33.3 A physician would like to determine the global pat- terns of gene expression in two different types of tumor cells in order to develop the most appropriate form of chemotherapy for each patient. Which of the following techniques would be most appropriate for this purpose? A. Southern blot. B. Northern blot. C. Western blot. D. ELISA. E. Microarray. 33.4 A 2-week-old infant is diagnosed with a urea cycle defect. Enzymic analysis showed no activity for ornithine transcarbamoylase (OTC). Molecular analy- sis revealed that the mRNA product of the gene for OTC was identical to that of a control. Which of the techniques listed below was most likely used to ana- lyze the size and amount of the mRNA? A. Dideoxy chain termination.B. Norther blot. C. Polymerase chain reaction. D. Southern blot E. Western blot Correct answer = E. Both the father and mother must be carriers for this disease. The son must have inherited a mutant allele from each parent. Because he shows only the 3-kb band on the Southern blot, the mutant allele for this disease must be linked to the 3-kb band for both parents. The normal allele must be linked to the 4-kb band in both parents. Because the daughter inherited the 4-kb band from both parents, she must be homozygous normal for the hex- osaminidase A gene.Correct answer = E. Microarray analysis allows the determination of mRNA production (thus, gene expression) from thousands of genes at once. A Northern blot only measures MRNA pro- duction from one gene at a time. Western blots and ELISA measure protein production (also gene expression), but only from one gene ata time. Southern blots are used to analyze DNA, not gene expression. Correct answer = B. Northern blot allows analy- sis of the mRNA present (expressed) in a partic- ular cell or tissue. Southern blot is used for DNA analysis, whereas Western blot is used for pro- tein analysis. Dideoxy chain termination is used to sequence DNA. Polymerase chain reaction (PCR) is used to generate multiple, identical copies of a DNA sequence in vitro.