HEMOGLOBIN

STRUCTURE & FUNCTION

By
SARANYA WWR
TUTOR
DEPARTMENT OF BIOCHEMISTRY
PSPMCH & RI
• Specific Learning Objective:

• By the end of this lecture, the students should

be able to know:

• The structure and function of hemoglobin.

• The factors affecting oxygen binding to hemoglobin.

• Examples of normal and abnormal hemoglobin

structures.
• Hemoglobin (Hb)
–A hemeglobin (heme + protein) found only in
red blood cells.
–Oxygen transport function.
–Contains heme as prosthetic group.
• Prosthetic means it acts as a co-factor
• (it doesn’t consume the oxygen itself but it
delivers O₂ to the tissues).
• Heme is present in
a. Hemoglobin
b. Myoglobin
c. Cytochromes
d. Peroxidase
e. Catalase
f. Tryptophan pyrrolase
g. Nitric oxide synthase
Hemoglobin functions :
 Carries oxygen from the lungs to tissues
 Carries carbon dioxide from tissues back to the
lungs
 Normal levels (g/dL):
Males: 14-16
Females: 13-15
Overview of lecture
 Structure
• It is conjugated protein having heme as the
prosthetic group and the protein is globin.
• It is tetrameric protein with 4 subunits, each subunit
having a prosthetic heme group and globin
polypeptide.
• The polypeptide chains are usually two alpha and
two beta chains.
• Alpha contain 141 amino acids & beta chain
contain 146 amino acids.
• Hemoglobin has a molecular weight of about
67,000 Daltons.
• Each gram of Hb contains 3.4 mg of iron.
• Heme is a derivative of the porphyrin.
• Porphyrins are cyclic compounds formed by
fusion of 4 pyrrole rings linked by methenyl
(=CH-) bridges.
• Since an atom of iron is present, heme is a
ferroprotoporphyrin.
• The pyrrole rings are named as I, II, III, IV
and the bridges as alpha, beta, gamma and
delta.
Types of hemoglobin in adults
Hemoglobin
Normal
HbA (90%)

HbA2 (2%)

HbF (1%)

HbA1c (6%)

Abnormal
Carboxy Hb

Met Hb

Sulf Hb
• Hemoglobin A (HbA)
– Major Hb in adults.

– Composed of four polypeptide chains:

 Two α two β chains.

– Contains two dimers of αβ subunits (each dimer has 2 heme)

– Held together by non-covalent (hydrophobic) interactions.

– Each chain is a subunit with a heme group in the center that

carries oxygen.

– A Hb molecule contains 4 heme groups and carries 4 molecules

of O₂
 Factors affecting O2 binding:
Factors that reduce the
ability of hemoglobin to
bind to O2 so it can be
easily delivered to the
tissues .
3 allosteric effectors:

pCO2
Po2
pH of the (partial Availabilit
(Partial
environme carbon y of 2,3-
oxygen
nt dioxide BPG
pressure)
pressure)
• Oxygen dissociation curve :
–The curve is sigmoidal in shape

–Indicates co-operation of subunits in O₂

binding

–Binding of O₂ to one heme group increases O₂

affinity of others

–Heme-heme interaction
 Factors affecting oxygen binding :

•1. pO₂ (partial oxygen pressure) :

– P50 (mm Hg): the pressure at which Hb is 50%
saturated with O₂
– Indicates affinity of Hb to O₂ :
A. High affinity → slow unloading of O₂ (low P50
value)
B. Low affinity → fast unloading of O₂ (high P50
value)
Lung pO₂ is 100 mm → Hb saturation 100%
Tissue pO₂ is 40 mm → Hb saturation reduces *
Hence O₂ is delivered to tissues
 • 2. pH of the environment

pH is the negative log of hydrogen ion concentration .

If the hydrogen ion concentration high the ph will be
low and if the hydrogen concentration low the ph will
be high

1- Decrease in pH = increased acidity

2- increasing carbon dioxide pressure
Are both important factors for oxygen delivery
•pCO₂ (partial carbon dioxide pressure) :

– The Bohr effect :

• It is the shift of the ODC to the right in response to an increase in pCO2 or a decrease in
pH
• Is the effect of pH and pCO₂ on:
A. Oxygenation of Hb in the lungs
B. Deoxygenation in tissues

– Tissues have lower pH (acidic) than lungs

 Due to proton generation (two reactions):
• CO₂ + H₂O H2CO₃
• H2CO₃ HCO₃⁻ + H+
– Protons reduce O2 affinity of Hb Causing easier O2 release into the tissues.
– The free Hb binds to two protons.
– Protons are released and react with HCO3 – to form CO₂ gas
• (HCO₃⁻ + H+  CO₂ + H₂O )
– The proton-poor Hb now has greater affinity for O₂ (in lungs).
– The Bohr effect removes insoluble CO₂ from blood stream and produces soluble bicarbonate
.
4. Availability of 2,3- BPG :
– Binds to deoxy-Hb and stabilizes the T-form.
– When oxygen binds to Hb, BPG is released.
Compensatory mechanism

At high altitudes and “hypoxia” :

 RBC number increases
 Hb concentration increases
 BPG** increases
 This decreases O2 affinity of Hb
 Thus increases O2 delivery to
tissues
 Factors affecting oxygen binding
• Oxygen affinity :
 High O2 affinity is due to:
1. Alkalosis
2. High levels of Hb F
3. Multiple transfusion of 2,3 DPG-depleted blood
• Abnormal hemoglobin :
• They are unable to transport O₂ due to
abnormal structure
Sulf-HB: Forms due to Carboxy-Hb: CO₂
high sulfur levels in replaces O₂ and binds Met-Hb: Contains
blood 220X tighter than O₂ oxidized Fe³⁺ (~2%)
(irreversible reaction) (in smokers) that cannot carry O₂
 Other hemoglobin forms
Fetal Hemoglobin (HbF) HbA2 HbA1c
Tetramer with two a and two g Composed of two a and two d Two α and two β -Glucose
chains globin chains

Major hemoglobin found in the HbA1c levels are high in

Appears shortly before birth.
fetus and newborn patients with diabetes mellitus

 Higher affinity for O2 than  Constitutes ~2% of total Hb  HbA undergoes non-
HBA enzymatic glycosylation
 Glycosylation depends on
plasma glucose levels
Important because the Because they have higher
maternal O2 which is mainly amount of glucose
HbA has to be delivered to the *life span of RBCs =120
fetal hemoglobin across the day so if the glucose
placenta which is HbF so it has bound to HbA1c level
because it stays for 120
tohave higher
Transfers affinity
O2 from to O2
maternal
to fetal circulation across days so until the patient
placenta maintain his blood glucose
level over a period of 3
months
*fasten glucose test will
show drop in glucose
because the patient didn’t
 Summary
Hb functions : Carries oxygen from the lungs to tissues .Carries carbon dioxide from
tissues back to the lungs
Normal levels : Males: 14-16, Females: 13-15
A.High affinity → slow unloading of O₂ (low P50 value)
B. Low affinity → fast unloading of O₂ (high P50 value)
pO₂ - Lung pO₂ is 100 mm → Hb saturation 100%
- Tissue pO₂ is 40 mm → Hb saturation reduces
Acidosis :Decreased O₂ affinity
pH
Alkalosis: increased O₂ affinity

The Bohr effect : Is the effect of pH and pCO₂ on

Factors affecting oxygen binding

A. Oxygenation of Hb in the lungs B. Deoxygenation in tissues.

-Tissues have lower pH (acidic) than lungs
CO₂ + H₂O → HCO₃⁻ + H⁺
pCO₂ -Protons reduce O2 affinity of Hb
-The free Hb binds to two protons
- Protons are released (HCO₃⁻ + H⁺ → CO₂ + H₂O )
-The proton-poorHb now has greater affinity for O₂ (in lungs)
-The Bohr effect removes insoluble CO₂ from blood stream and produces soluble bicarbonate .

Availability of -Binds to deoxy-Hb and stabilizes the T-form

2,3- -When oxygen binds to Hb, BPG is released
bisphosphogly At high altitudes and “hypoxia” BPG increases
cerate This decreases O2 affinity of Hb , Thus increases O2 delivery to tissues
THANK YOU