DR.

IMRAN ALI
(BSc,MBBS,MPHIL)
Assistant Professor
Biochemistry FMH
HEMOGLOBIN
ALLOSTERIC??
OXYGEN HEMOGLOBIN DISSOCIATION CURVE

DEFINITION:
CURVE INDICATES THAT OXYGEN BINDING BY Hb
DEPENDS ON PARTIAL PRESSURE OF OXYGEN
T STATE/R STATE
Factors Affecting Oxygen-hemoglobin
Dissociation Curve
Oxygen
loading
Oxygen-hemoglobin dissociation curve is shifted to right in the
following conditions:
1) Decrease in partial pressure of oxygen
2)1) Increase
Decrease inin partial
partial pressure
pressure ofof oxygen
carbon dioxide (Bohr effect)
2) Increase in partial pressure of carbon dioxide (Bohr effect)
3) Increase in hydrogen ion concentration and decrease in pH
3) Excess of 2,3-diphosphoglycerate (DPG) in RBC.
(acidity)
4) Increase in hydrogen ion concentration and decrease in pH
4) Increased
(acidity) body temperature
5)5) Excess of 2,3-diphosphoglycerate
Increased body temperature (DPG) in RBC.

(It is also called 2,3-biphosphoglycerate (BPG). DPG is a byproduct in glycolysis. It

combines with β-chains of hemoglobin. In conditions like muscular exercise and in
high altitude, the DPG increases in RBC. So, the oxygen-hemoglobin dissociation
curve shifts to right to a great extent)

Shift to right:
 2,3 BPG

Fetal Hb (HbF) – binding ability of BPG to HbF is LOW .

This facilitates trans-placental O2 transfer
 BOHR EFFECT

• “The binding of oxygen to hemoglobin

decreases with increasing H+ concentration
(low pH) or when the hemoglobin is exposed
to increased partial pressure of CO2 (pCO2)”
• This phenomenon is known as Bohr effect.
• lt is due to a change in the binding affinity of
oxygen to hemoglobin.
• Bohr effect causes a shift in the oxygen
dissociation curve to the right
 EFFECT OF TEMP. ON ODC
When cells have a high metabolic rate, they produce an excess amount of
thermal energy as a waste by-product.

This thermal energy when transferred to blood via process of heat, it increases
the average kinetic energy of the molecules and particles within the plasma,
thereby increasing its temperature.

A higher temperature is correlated to the cells working harder and therefore

means they need a higher supply of oxygen to keep them going.

Therefore at higher blood plasma temperatures, the hemoglobin becomes less

likely to bind to oxygen and much more likely to unload OXYGEN into the cells
of the tissue.

Therefore, as temperature increases, this shifts the entire oxygen-

hemoglobin dissociation curve to the right.

This ultimately means that the exercising cells will receive more
oxygen
CARBON MONOXIDE EFFECT ON ODC
“CADET” face right
HEMOGLOBIN BREAKDOWN
HEMOGLOBIN
OXYGENASE

BILIVERDIN
REDUCTASE
 SOME IMPORTANT HEMOPROTEINS
• HEMOPROTEINS ARE THE PROTEINS WHICH CONTAIN HEME AS TIGHTLY
BOUND PROSTHETIC GROUP .
Heme
a
(TYPES OF JAUNDICE)
Hyperbilirubinemia (jaundice)
 2. Hepatic:
1) Gilbert's disease
• May be caused by inability of the hepatocytes to uptake bilirubin
from the blood
 Associated with reduced activity of Glucuronyl-transferase enzyme.
• As a result, unconjugated bilirubin accumulates.
2) Physiological jaundice
3) Crigler-Najjar syndrome
• Conjugation is impaired due to severely defected Glucuronyl
transferase enzyme
• Unconjugated bilirubin is increased in the body.

4) Dubin-Johnson syndrome
Inability of hepatocytes to secrete conjugated bilirubin after it has
been formed.
• Conjugated bilirubin returns to the blood.
 3. Post-hepatic (biliary obstruction)

• e.g., biliary calculi causes backup (interference

with the secretion) and reabsorption of
conjugated bilirubin. Blood levels of
conjugated bilirubin increase.
Genetic disorders of Bilirubin
metabolism
CLINICAL DISORDERS OF
HEMOGLOBIN
ABNORMALITIES
 EXAMPLES OF HEMOGLOBINOPATHIES
• Sickle cell anemia (Hb S),
• Hemoglobin C disease (Hb C),
• Hemoglobin SC disease (Hb S + Hb C)
• Thalassemia

 The first three conditions result from production of hemoglobin

with an altered amino acid sequence (qualitative
Hemoglobinopathy),
 Whereas the thalassemias are caused by decreased production of
normal hemoglobin (quantitative Hemoglobinopathy).
 Sickle cell anemia
• Sickle cell anemia is a genetic disorder of the blood
caused by mutation in the gene for β-globin chain of
hemoglobin.
• The mutant β-globin chain is designated as βS, and
the resulting hemoglobin(α2β2S) is referred to as
HbS.
 Sickle cell anemia
• A molecule of Hb S contains two normal α-globin chains and
two mutant β-globin chains (βS), in which glutamate at
position 6 has been replaced by valine.
 SICKLE CELL ANEMIA:
Sickle cellPICTURE
CLINICAL anemia
 At low oxygen tension, Hemoglobin S has the tendency to
polymerize inside the RBC, forming a network of fibrous polymers
that stiffen and distort the cell, producing rigid, misshapen and
sickle shaped RBCs. Such sickled cells frequently block the flow of
blood in the narrow capillaries.
 This interruption in the supply of oxygen leads to localized anoxia
(oxygen deprivation) in the tissue, causing pain and eventually
death (infarction) of cells in the vicinity of the blockage

VALINE IS INSOLUBLE AS ITS NONPOLAR.

GLUTAMATE IS SOLUBLE AS ITS POLAR.
 THINK…

HBS
 Sickle cell anemia
• Sickle cell anemia is characterized by lifelong
episodes of pain (“crises”), chronic hemolytic
anemia with associated hyperbilirubinemia.
• Increased susceptibility to infections, usually
beginning in early childhood.
SICKLE CELL ANEMIA:
TREATMENT
 Hemoglobin C disease
• Like Hb S, Hb C is a hemoglobin variant that has a single
amino acid substitution in the sixth position of the β-globin
chain.
• In this case, however, a lysine is substituted for the
glutamate (as compared with a valine substitution in Hb S).

• Patients of hemoglobin C generally have a relatively mild,

chronic hemolytic anemia. These patients do not suffer from
infarctive crises, and no specific therapy is required
Hemoglobin SC disease
Before starting thalassemia
Lets first discuss
Hemoglobin gene family
• HEMOGLOBIN GENES DIRECT THE SYNTHESIS OF HEMOGLOBIN
CHAINS
• The genes coding for the α-globin-like and β-globin-like subunits of the hemoglobin chains
occur in two separate gene clusters (or families) located on two different chromosomes.
• α--Gene family :The α-gene cluster on chromosome 16 contains two genes for two α-
globin chains α1 and α2 . It also contains the ζ gene …mainly expressed in embryonic life.
• β-Gene family: A single gene for the β-globin chain is located on chromosome 11. There
are an additional four β-globin-like genes:
 Thalassemias
• The thalassemias are hereditary hemolytic diseases
in which an imbalance occurs in the synthesis of
globin chains.

• Normally, synthesis of the α- and β-globin chains is

coordinated, so that each α-globin chain has a β-
globin chain partner. This leads to the formation of
α2β2 (Hb A).

• In the thalassemias, the synthesis of either the α- or

the β-globin chain is defective.
 Classification of Thalassemia
(1) Each thalassemia can be classified as either a
disorder in which no globin chains are produced:
(αo- or βo-thalassemia)
(2) Or one in which some chains are synthesized, but
at a reduced level:
(α+-thalassemia or β+-thalassemia)
 β-Thalassemias:
• In these disorders, synthesis of β-globin chains is decreased or
absent, typically as a result of gene mutations that direct the
synthesis of β-globin chains; however, α-globin chain synthesis is
normal.
• α-Globin chains cannot form stable tetramers and, therefore,
precipitate, causing the premature death of cells initially destined
to become mature RBCs.
• Increase in α2γ2 (Hb F) and α2δ2 (Hb A2) also occurs.
β-Thalassemias:
 Treatment of β-thalassemia
• β-thalassemia minor:
• Those individuals with β-thalassemia minor make some β
chains, and usually do not require specific treatment. Can
spend a normal life.

• β-thalassemia major:
• These patients require regular transfusions of blood.
• However excessive blood transfusion can cause iron
overload (Haemosiderosis)
• For excessive iron load, use of iron chelation therapy is
needed.
 α-Thalassemias:
• These are defects in which the synthesis of α-globin
chains is decreased or absent, typically as a result of
genetic mutations.
• Because each individual’s genome contains four copies
of the α-globin gene (two on each chromosome
16),there are several levels of α-globin chain
deficiencies as follows;
• (1) SILENT CARRIER:
• If one of the four genes is defective, the individual is termed
as silent carrier of α-thalassemia, because no physical
manifestations of the disease occur.
• (2) α- THALASSEMIA TRAIT:
• If two α-globin genes are defective, the individual is
designated as having α –thalassemia trait.
 α-Thalassemias:
• HB H DISEASE :
• If three α-globin genes are defective, the individual has Hb
H disease. Lives upto late childhood or early adulthood

• HB BART (γ4) : (ALPHA THALASSEMIA MAJOR)

• If all four α-globin genes are defective,
• Hb Bart (γ4) disease with hydrops fetalis and fetal death
results
• Because α-globin chains are required for the synthesis of
Hb F.
• So fetus cannot survive.