e d

e r
A presentation on
s t
B.Sc. Semester - II, Core Course – IV: ZOO 202 – C,
g i
Animal Physiology, Endocrinology : Unit – 4(2)
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53
STRUCTURE AND FUNCTION OF
.
r
HEMOGLOBIN 5
r t e
v e
o n Dr O. Hemchandra
Assistant Professor
C
F
Department of Zoology

D Kumbi College, Kumbi

P
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Hemoglobin (Hb)
e r
• A hemeprotein found only in red blood cells
s t
• Oxygen transport function
g i
• Contains heme as prosthetic group
r e
n
• Heme reversibly binds to oxygen (the point of reversible
U
binding is whenever the oxygen is high it takes it and
5 3
whenever it is low it releases it)
5 .
e r
Prosthetic group: Helper molecule that is bound to a protein

r t
The heme group
v e
o n
• A complex of protoporphyrin IX and ferrous iron (Fe+2)

• Fe+2 binds toF

C
• Fe+2 is present in the center of the heme
four nitrogen atoms of the porphyrin ring
• Forms two D additional bonds with:
• HistidineP
b
• Oxygen
e residue of globin chain
molecule not atom.
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 e d
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s t
*Heme group has iron in
g i
the center in ferrous
r e
state , if it is in ferric
n
state it will not bind to U
oxygen
5 3
*in ferrous state it can
5 .
have 6 bonds 4 with
e r
nitrogen we call it r t
porphyrin ring 1 bond
v e
with molecular O2
o n
(molecule (O2) NOTE C
F
atom (O)) 1 attached to
D
the
P
globin chain

e
(polypeptide)
b
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Types of hemoglobin in adults
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HbA (90%)
s t
g i
e
HbA2 (2%)

n r
U
Normal HbF (1%)

5 3
HbA1c (6%)

5 .
Hemoglobin
e r
r t Carboxy Hb

v e
Abnormal
o n Met Hb

C
F
Sulf Hb

D
P
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Hemoglobin A (HbA)
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• Major Hb in adults
e r
• Composed of four polypeptide chains:
s t
i
1 dimer = 2 subunits

g
* Two α and two β chains
( thus Hb composed
• Contains two dimers of αβ subunits
r e
of 4 subunits)

n
• The dimers are held together by non-covalent

U
interactions
• Each chain is a subunit with a heme group in the
center that carries oxygen
5 3
• A Hb molecule contains 4 heme groups and carries
5 .
4 molecules of O2

e r
r t
v e
o n Between α and β there are
C hydrogen, Ionic and
F hydrophobic bonds while
D between the two dimers
P there are ionic and

b e hydrogen bonds. (α and β

d o are held together stronger

than the two dimers)

A
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HbA Structure
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5 3
5 .
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C
F
D
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T-form of Hb
s t
• The deoxy form of Hb
g i
• Taut form (or tense form)
r e
• The movement of dimers is constrained
n
• Low-oxygen-affinity form
U
5 3
R-form of Hb
5 .
• The oxygenated form of Hb

e r
• Relaxed form
r t
e
• The dimers have more freedom of movement
• High-oxygen-affinity form
n v
o
C
F
When Hb reaches the tissue, it becomes more

D
taut so O2 will be released (like squeezing).

P
While in lungs, Hb will be in relaxed form in

e
which more O2 will be able to be bound.
b
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Hemoglobin function
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• Carries oxygen from the lungs to tissues
e r
• Carries Carbon dioxide from tissues back to the lungs
s t
• Normal level (g/dL):
g i
•
•
Males: 14-16
Females: 13-15 r e
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Factors affecting oxygen binding

5 3
•
•
Three allosteric effectors:
pO2 (Partial oxygen pressure)
5 .
• pH of the environment
e r
• pCO2 (partial carbon dioxide pressure)
r t
• Availability of 2,3-bisphosphoglycerate
v e Hint: BPG = DPG

Oxygen Dissociation curve o n

• C
The curve is sigmoidal (sigmoidal = S-shaped)
•
F
Indicates cooperation of subunits in O2 binding As we know, we have 4
•
D
(cooperation (cooperative binding): when the oxygen subunits, so when 1
P
binds to hemoglobin it helps the binding of the next subunit binds to O2, the
oxygen)
b e other subunits will bind

o
• Binding of O2 to one heme group increases O2
to O2 also.

d
affinity of others Heme-heme interaction

A
 e d
pO2 (partial oxygen pressure)
e r
s t
• Indicates affinity of Hb to O2
g i
• P50(mmHg): the pressure at which Hb is
r e
50% saturated with O2
n
If P50 is high = low
• High affinity → slow unloading of O2 U affinity

53
(occur in lungs) Cause: we need

• Low affinity → fast unloading of O2

5 . more pressure to
saturate O2 within
(occur in tissues)
e r RBCs.
• Lung pO2 is 100 mm → Hb saturation
r t &
100%
v e Vice versa

• n
Tissue pO2 is 40 mm → Hb saturation
o
reduces
C
•
F
Hence O2 is delivered to tissues
D
P
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5 3
5 .
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r t
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C
F
D
P
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The Bohr effect
e r
• Effect of pH and pCO2 on:
Bohr : scientist name

s t
• Oxygenation of Hb in the lungs (pH is high in
g i
e
the lung)
• Deoxygenation in tissues (pH is low in the
tissues) n r
• Tissues have lower pH (acidic) than lungs U
• Due to proton generation:
5 3
• CO2 + H20  HCO3- + H+ (H+ is more than
5 .
r
HCO3- , So the environment will be acidic)
•
•
Protons reduce O2 affinity of Hb

r
Causing easier O2 release into the tissuest e
• The free Hb binds to two protons
v e
•
n
Protons are released and react with HCO3 –
to form CO2 gas
o
•
C
The proton-poor Hb now has greater affinity
• for O2 (in lungs)
F
•
D
The Bohr effect removes insoluble CO2( in

P
the form of HCO3-) from blood stream
•
e
Produces soluble bicarbonate

b
d o
A
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Proton generation in the tissue has many sources, thus the
e r
net result will be acidic pH in tissue despite HCO3 -
s t
generation.
g i
• If the curve shifts to the left (higher PH) = higher affinity; r e
less pressure is required for saturation. n
U
3
• If the curve shifts to the right (lower PH) = lower affinity;
5
more pressure is required for saturation.
5 .
• In lungs, pH is high (Alkaline), thus the affinity is high for
e r
O2 binding.
r t
v e
o n
Availability of 2,3 bisphosphoglycerate
C
(one of the glycolysis products)
F
• Binds to deoxy-hb and stabilizes the T-form (it reduces
D
the affinity)
P
e
• When oxygen binds to Hb, BPG is released
b
d o
A
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At high altitudes:
e r
• RBC number increases
s t
• Hb conc. Increases
g i
•
e
BPG increases → reduces the
r
n
affinity, why? To ensure O2

U
delivery to the tissues.

5 3
5 .
e r
r t
v e
o n
C
F
D
P
b e
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A
 e d
e r
s t
High altitude and O2 affinity
g i
In hypoxia and high altitude
r e
• 2,3 BPG levels rise n
• This decreases O2 affinity of Hb U
• Thus increases O2 delivery to tissues
5 3
5 .
e r
r t
BPG: is produced by tissues

v e
BPG is not bound to Hb when it
n
reaches the lungs, but once Hb
o
C
reaches to the tissue,

F BPG will bind to it causing release

D of O2
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High O2 affinity is due to:
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• Alkalosis
e r
s t
i
• High levels of Hb F
• Multiple transfusion of 2,3 DPG-depleted blood
e g
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U
5 3
5 .
e r
r t
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o n
C
F
D
P
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A
Fetal hemoglobin (HbF)
e d
• Major hemoglobin found in the fetus and
e r
s t
i
newborn
• Tetramer with two  and two  chains
e g
•
•
Higher affinity for O2 than HBA
Transfers O2 from maternal to fetal n r
circulation across placenta U
HbA2
5 3
• Appears ~12 weeks after birth
5 .
• Constitutes ~2% of total Hb

e r
• Composed of two  and two δ globin chains
r t
e
HbA1c
• HbA undergoes non-enzymatic
glycosylation n v
o
•
C
Glycosylation depends on plasma glucose
levels
F
•
D
HbA1c levels are high in patients with

P
diabetes mellitus

b e
d o
HbA1c has a half life of 2-3 weeks, so you can tell about the blood glucose
level of 3 weeks ago.
A
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Abnormal Hbs
n
Unable to transport O2 due to abnormal structure U
• Carboxy-Hb: CO replaces O2 and binds 200X (200 times)
5 3
tighter than O2 (in
5 .
• smokers) (reversible)
e r
r t
• Met-Hb: Contains oxidized Fe3+ (~2%) that cannot carry
O2 (reversible)
v e
o n
• Sulf-HB: Forms due to high sulfur levels in blood
(irreversible reaction) C
F
D
P
b e
d o
A