Metabolism of

Nucleoproteins

Q.1. What are the sources of nitrogen and carbon atoms of the
purine ring?
Sources:
• N1 = amino nitrogen of
aspartate
• C2 and C8 = For–mate via
F.H4 or β carbon of ser-
ine/or glycine
• N3 and N9 = Amide N2 of
glutamine
• C4, C5 and N7 = From
Glycine
• C6 = from CO2
Q.2. What are the sources of the nitrogen and carbon atoms of
the pyrimidine ring?
Sources:
• C2 = from CO2
• N3 = from amide N of
glutamine
• N1 and C4, 5 and 6 = from
aspartic acid.

Q.3. What are the ingredients required for the synthesis of

pyrimidines?
Ingredients required are:
• Carbamoyl phosphate: Synthesized from CO2 and amide
N of glutamine.
• PRPP: 5-phosphoribosyl-1-pyrophophate
 • Enzymes: Carbamonyl-P synthetase II (cytosolic enzyme),
– Transcarbamoylase
– Dihydro-orotase
– Dehydrogenase
– Transferease
– Decarboxylase
• ATP: For energy
• Amino acid: Aspartic acid
• Coenzymes/cofactors: FAD, NAD+, Mg++
Q.4. What is orotic acid?
• It is an intermediate metabolite in pyrimidine synthesis.

Q.5. What is the first Pyrimidine nucleotide formed?

• Uridylic acid (UMP) is the first pyrimidine nucleotide
formed.
• Other pyrimidine nucleotides viz. UDP, UTP, CTP and
d-UDP are synthesized from UMP.
Q.6. What is orotic aciduria?
An inherited disorder of pyrimidine synthesis. Two types:
• Type I orotic aciduria
– Enzymes defect: orotate phosphoribosyl transferase
and orotate decarboxylase.
– Orotic acid is not converted to UMP. Results in
accumulation of orotic acid in blood increasing its
level. There is growth retardation and megaloblastic
anemia.
• Type II orotic aciduira:
– Enzyme defect: Orotidylate decarboxylase. It is also
characterized by megaloblastic anemia and urinary
excretion of orotic acid.
Q.7. State the end products of pyrimidine catabolism. Where
pyrimidine catabolism takes place?
• Liver is the main site of catabolism of pyrimidines.
 • End products of catabolism are:
– CO2 and NH3—end products
– β-alanine-side product from cytosine and uracil.
– β-aminoisobutyrate: Side product formed from
catabolism of thymine, it is excreted in urine.
Q.8. What is the fate of β-alanine formed in pyrimidine
catabolism?
• β-alanine can be utilized for synthesis of
– CoA-SH
– Carnosine
– Anserine
• Alternatively, β-alanine can be oxidized to acetate, NH3
and CO2.
Q.9. What is the clinical significance of β -amino-isobutyric
acid?
• β-amino isobutyric acid is excreted in large quantities in:
– Leukemias and
– When body is subjected to X-ray irradiation (β β-amino
isobutyric aciduria)
• β-amino isobutyrate can be converted to methyl malonic
semialdehyde which, in turn, can form propionic acid
which is converted to succinate (Thus it is glucogenic.).
Q.10. State the materials required for purine biosynthesis.
• PRPP (5-phosphoribosyl-1-pryophosphate): is the
starting material. It is formed from D-ribose-5'-P
obtained from HMP shunt.
• Enzymes: Various enzymes synthetases, transferases,
carboxylase, and hydroxylases.
• Engery: Provided by ATP.
• Amino acids and derivatives: Glycine, aspartic acid and
glutamine.
• CO2: from HCO3–
• Coenzymes/and cofactors: F.H4, Mg.++
Q.11. Name the reaction which incorporates CO2 to the
substrate but it is not “CO2-fixation reaction”.
• Formation of 5-amino-imidazole-4-carboxylic acid
ribotide (C-AIR)
 • The reaction uses CO2 to carboxylate 5-amino-imidazole
ribotide (AIR). It contributes to C6 of the purine nucleus.
• The reaction is peculiar in that neither biotin nor ATP is
required for this carboxylation reaction.
Q.12. What is the first purine nucleotide formed in purine
biosynthesis?
• First purine nucleotide formed is inosine monophosphate
(IMP).
• Once IMP is formed, it can form other purine nucleotides
viz. AMP and GMP.
Q.13. How PRPP is formed?
• PRPP is formed from ribose-5-P and ATP catalyzed by
the enzyme PRPP synthetase.

Q.14. What are various uses of PRPP?

PRPP is used for:
• Synthesis of both purine and pyrimidine nucleotides.
• Salvage pathways for both purine and pyrimidine
bases.
• Biosynthesis of nucleotide coenzymes.
Q.15. How many high energy phosphate bonds are utilized in
purine synthesis?
Expenditure of six high energy phosphate bonds are
utilized in purine synthesis, thus it is energetically an
expensive process.
Q.16. How AMP is formed from IMP?
• Aspartic acid condenses with IMP to form adenylo-
succinate, catalyzed by the enzyme “adenylosuccinate
synthetase” and Mg++. GTP provides the energy.
• Adenylosuccinate is then cleaved to form fumaric acid
and AMP.
Q.17. How GMP is formed form IMP?
• IMP is first oxidized to xanthylic acid (xanthine
monophosphate, XMP).
• Oxidation is catalyzed by a dehydrogenase, NAD+ acting
as H-acceptor.
• Glutamine gives the amide group to C2 of XMP to form
GMP.

Q.18. What is meant by salvage pathways for purine/

pyrimidine bases?
Many cells have pathways that can “salvage” the purine
and pyrimidine bases to form the corresponding
nucleotides and do not require expenditure of energy. Such
pathways are called “salvage” pathways.
Q.19. What are the salvage pathways by which purine bases
form nucleotides?
Two pathways are available for formation of nucleotides by
salvage pathway from purine bases. They are:
• One step synthesis
• Two step synthesis.
Q.20. State one step synthesis of purine nucleotides by salvage
pathway.
a. Formation of GMP and IMP:
• “Hypoxanthine-Guanine phosphoribosyl transferase”
(HGPRTase) enzyme catalyzes one-setp formation of
the nucleotides from either guanine or hypoxanthine,
using PRPP as the donor of ribosyl moiety.
 b. Formation of AMP:
• The enzyme “adenine phosphoribosyl transferase”
(APRTase) catalyzes the formation of AMP from
adenine, ribosyl moiety is donated by PRPP.

Q.21. State the two-step synthesis of AMP?

• Also called as nucleoside phosphorylase-nucleoside
kinase pathway.

Q.22. Can GMP/IMP be formed by two-step synthesis?

Neither Guanosine nor inosine kinase have been detected
in mammalian cells. So GMP/IMP cannot be formed by this
pathway. Adenine is the only purine base that can be
salvaged by two-step pathway.
Q.23. Which tissues cannot synthesize the purine nucleo-
tides?
Erythrocytes, neutrophils and brain cells are not capable of
“de Novo” synthesis of purine nucleotides as they lack the
enzyme “PRPP-amidotransferase”.
Q.24. What is the end-product of catabolism of purines in
humans?
Uric acid is the chief end-product of purine catabolism in
humans and primates.
Q.25. Show schematically how uric acid is formed in humans.

Q.26. Name the enzymes of uric acid metabolism which are

absent in humans but present in lower animals. What is
their function?
• Enzymes: uricase and allantoinase are absent in humans.
• In many non-primates, uric acid may be oxidized and
decarboxylated by uricase, hepatic-Cu containing
enzyme to form allantoin.
• Some fishes have uricase, as well as allantoinase which
converts allantoin to allantoic acid.

Q.27. What is the nature of the enzyme xanthine oxidase?

• Xanthine oxidase is a metallo-enzyme and contains
molybdenum (Mo), the trace element and also Fe3+.
• It can act on hypoxanthine and converts it to xanthine.
Also it can act on xanthine and converts it to uric acid.
• The enzyme requires FAD and molecular O2 and
produces H2O2 at substrate level.
Q.28. What is the normal uric acid level of blood?
• In males: 3.0 to 8.5 mg/dl.
• In females: 2.0 to 7.5 mg/dl.
Q.29. How much uric acid is excreted daily in urine?
• 400 to 600 mg/24 hrs urine.
Q.30. Which is the main organ where uric acid formation take
place?
Liver is the principal organ. After formation uric acid is
carried to kidney for its excretion.
Q.31. What is meant by miscible pool?
It is the quantity of uric acid present in body water. In a
normal adult, an average of 1130 mg of uric acid is
present.
Q.32. What is meant by the turnover of uric acid?
This indicates the rate at which the uric acid is synthesized
and lost from the body. Normally 500 to 600 mg of uric acid
is synthesized. Not all is excreted in urine, some uric acid is
secreted in bile to reach the gut.
Q.33. What happens to uric acid in the gut secreted in bile?
By the action of intestinal bacteria, uric acid is converted to
urea and NH3.
Q.34. How uric acid is excreted?
• Uric acid in plasma is filtered by the glomeruli but is
later partially reasorbed by the renal tubules.
• There is also now conclusive evidence for tubular
secretion of uric acid by kidney.
Q.35. State the effects of hormones on uric acid excretion.
• Gluco-corticoids and
• ACTH increases the excretion of uric acid in urine.
Q.36. What are uricosuric drugs. Name them.
Uricosuric drugs are those that block reabsorption of uric
acid by the renal tubules thus enhancing the excretion of
uric acid in urine.
Examples:
• Salicylates
• Probenecid (Benemide)
• Halofenate
Q.37. Name the substances that can competitively inhibit the
uric acid excretion.
Increased Lactic acid in blood as in Lactic acidosis
competes with uric acid excretion resulting to retention of
uric acid in blood thus increasing blood uric acid level.
Q.38. What is hyperuricemia?
Increase of blood uric acid level above normal is called
hyperuricemia.
Q.39. What is hypouricemia? How it can happen and what are
the effects?
• Decrease of blood uric acid level below normal is called
hypouricemia.
• It can occur due to deficiency of xanthine oxidase so that
uric acid is not formed, there is increased excretion of
hypoxanthine and xanthine, due either genetic defect or
to severe liver damage.
• In severe xanthine oxidase deficiency, patient may
exihibit xanthinuria and xanthine lithiasis.
Note: Also can be produced by administering allo-purinol,
a xanthine oxidase inhibitor.
Q.40. What do you mean by tophi?
Tophi are elevated nodules appearing in periarticular
tissues of joints and cartilages of the ears. These are formed
due to deposition of crystals of sodium urate in the articular
and periarticular tissues of the joints.
Q.41. What is gout? What are the features?
Gout is a relatively rare chronic disease characterized by
recurrent attacks of acute pain and swelling at first
affecting only one joint, usually metatarso phalangeal joint
of the big toe, later becoming polyarticular. The features are:
• Excess of uric acid in the blood (hyperuricemia).
• Deposition of sodium monourate crystals in periarti-
cular tissues of joints and cartilages of ears producing
“tophi”.
• Recurring attacks of acute arthritis with swelling of joints.
Q.42. What are the types of Gout?
Two types:
• Primary gout
 – Primary metabolic gout
– Primary renal gout
• Secondary gout:
– Increase in purine catabolism viz. in leukemias,
polycythemia, prolonged fasting.
– Secondary renal gout: in renal failure.
– In von Gierke disease (Type 1 GSD).
Q.43. What is the biochemical cause of gout in von Gierke
disease (Type 1 GSD) ?
• Increased synthesis of purines followed by cata-
bolism.
• Increased lactic acid in blood (lactic acidosis) competes
with uric acid for excretion resulting to retention of uric
acid, hyperuricemia and gout.
Q.44. How the blood uric acid level can be lowered in gout?
• By giving purine free diets.
• By increasing renal excretion of uric acid by suitable
uricosuric drug like probenecid.
• By decreasing the synthesis of uric acid using xanthine
oxidase inhibitor e.g. allopurinol (zyloprin) drug.
• By increasing oxidation of uric acid by administration of
uric acid oxidase.
Q.45. How does allopurinol acts?
• Allopurinol has similar structure like hypoxanthine.
Thus allopurinol acts by competitive inhibition of the
enzyme xanthine oxidase and thus uric acid synthesis is
inhibited.
• The durg causes a rapid fall in serum uric acid level.
Q.46. Outline briefly the line of treatment of a case of Gout?
Treatment of gout consists of:
• Palliative treatment
• Specific treatment
• Palliative treatment:
– Bed rest in acute stage.
– Diet: Purine free diet.
 – Anti-inflammatory drugs to relieve pain. Drugs used
are:
- Colchicine in acute attack.
- Non-steroid anti-inflammatory drugs (NSAIDs)
e.g. Indomethacin, diclophenin, naproxen, brufen,
pyroxicam, etc.
• Specific Treatment
Aims at lowering blood uric acid level.
– Xanthine oxidase inhibitor: Allopurinol (zyloprin)
drug of choice.
– Using uricosuric drugs by enhancing renal excre-
tion of uric acid e.g. probenecid (Benemide), halo-
fenate.
– By using drug to increase uric acid oxidations, e.g.
urate oxidase.
Q.47. What is Lesch-Nyhan syndrome?
• It is an X-linked recessive defect of the enzyme hypox-
anthine-guanine phosphoribosyl transferase (HGPRTase).
• Only males are affected.
• The enzyme is almost absent and leads to increased
purine salvage pathway from PRPP.
• Clinically manifests as severe gout, poor growth,
spasticity, tendency to self mutilation and renal failure.
Q.48. What is murexide test?
• Murexide test is used to detect uric acid in urine.
• To 5 to 8 drops of urine taken in a porcelain dish, 2 to 4
drops of conc. HNO3 is added. Heat carefully till every
trace of HNO3 and H2O removed. A reddish deposit of
purpuric acid is formed.
• Dip a rod in dilute NH3 and draw through the deposit.
It turns reddish-violet due to formation of ammonium
purpurate. Add to the same deposit a drop of 5% NaOH,
the deposit turns bluish-violet due to formation of Na-
purpurate.
Q.49. State the chemistry of Murexide test.
The uric acid is oxidized to dialuric acid and alloxan. These
two substances condense to form alloxantin. This
alloxantin so formed reacts with ammonium hydroxide to
form purpuric acid. The reddish-purple color is due to
formation of ammonium purpurate (Murexide).