Structure & Synthesis of haemoglobin (Hb)

Each Hb molecule is made up of:

• 4 Haem/Heme: Protoporphyrin (Pyrrol ring) + Fe

• Four Globin chains

 Haemoglobin (Hb)

Each of the 4 polypeptide chains is attached

to a haem moiety.

• For example, the normal adult Hb (Hb A)

has 2 alpha (a) & 2 beta (b) chains held
together by non-covalent bonds.

• Hb A = a2 b2
 Synthesis of Haemoglobin
• Haem & globin are produced at two different sites in the cell

• Haem in the mitochondria

• Globin in ribosomes

• Production of the 2 components of Hb occur simultaneously.

 Synthesis of Heme
• The heme moiety is made up of a protoporphyrin ring with an iron
atom attached at the centre.

• The main site of synthesis is the mitochondria

• Mature red cells do not have mitochondria

 Heme
synthesis
• Heme = Protoporphyrin + Iron
• Protoporphyrin = Iron free Heme
• Protoporphyrin consist of 4 pyrrol rings.
• Iron has 6 coordinations, 4 binds with the 4 pyrrol rings of
protoporphyrin, 1 binds with histidine (amino acid) from
globin chain and 1 is free (bind to Oxygen, CO 2 respectively)
Structure of Heme
1) Heme synthesis start with 2 simple molecules in the mitochondria

a. Glycine (amino acid)

b. Succinyl Co-A (intermediate of citric acid cycle)

2) Reaction of these 2 simple molecules with a co-factor (vitamin B6) will

produce aminolevulinic acid (ALA Synthase)
• This is the regulatory or rate limiting step of heme synthesis
3. ALA exits the mitocondria into the cytoplasm where some
steps occur to generate copropophyrinogen which enters back
into the mitochondria to form protoporphyrinogen and then
protoporphyrin.

4. Protoporphyrin bind with iron (ferrous- Fe2+)

At same time, globin is synthesized in the RER.

Protoporphyrin + ferrous + globin chains will form haemoglobin.
Synthesis of Haemoglobin
• When O2 binds to iron, the process is called oxygenation
not oxidation as oxygen is transported in circulation as a
molecule (O2).

• The oxidation state of iron rarely change. Oxidation of

Fe2+ to Fe3+ occur only occasionally. The oxidized form,
methemoglobin (1–2%), cannot bind to O2.
Synthesis of globin
 Types of Haemoglobin (Normal)
• Different types of Hb exist due to the type of globin
chain present & NOT Heme.

• Adults have and a small amount of its sub-type

HbA HbA2.

• Fetuses have HbF.

 Normal haemoglobin types
• Fetal Hb: HbF (a2g2)

• Adult Hb: HbA (a2b2)

• Adult Hb: HbA2 ( a2d2)

 HbF has more affinity for oxygen than HbA.

 Normal Adult Haemoglobin
Hb A Hb A2 Hb F

Structure a2b2 a2d2 a2g2

Normal % 96-98 % 1.5-3.2 % 0.5-0.8 %

 Abnormal haemoglobin
types
Qualitative defect in Hb
Hb S: caused by a substitution of amino acid in β globin chain
(valine for Glutamate).
Hb C: caused by a substitution of amino acid in β globin chain
(Lysine for Glutamate).
Others: Hb D, E, G, H to T

Quantitative defect in Hb (Thalasemias)

Major: β- thalassemia major (absence of synthesis of β globin chain-
no HbA (HbA2 & Hb F compensates).
Minor: reduction in synthesis of globin chain (α or β)
Normal & Abnormal
Haemoglobin
 Gas transport by haemoglobin
• Why is haemoglobin used as a transporter for gases? Because of the low
solubility of oxygen & CO2 in fluid. Therefore, haemoglobin has a
regulatory effect on oxygen concentration in the lungs and tissues.
Allosteric effect
• When a molecule reacts with conformational changes that increases or
reduces its activity, it is said to show an allosteric behaviour.
• Some substances act on Hb as allosteric substances.
• Although Hb is NOT an enzyme, it exhibit this characteristic of enzymes.
Substances that have an allosteric effect on Hb are:

1. 2,3 DGP (intermediate factor in glycolysis)

2. H+ ( i.e. pH)
3. CO2
 Forms of
Hb
• When Hb show high affinity to oxygen in the lungs it is in the
oxyhaemoglobin form refered to as the R form (R from Relax)
• When Hb show low affinity for oxygen in the tissues, it is in the
deoxyhaemoglobin form, refered to as T form (T from Tense)

• Allosteric substances act as stabilizers for T form of Hb i.e. they

affect the saturation of Hb with oxygen.
 Steps for O2- CO2
Transport
• In the lungs, the pressure of oxygen is high so it diffuses and
binds with Hb (bond of one O2 to haemoglobin fascilitates
the binding of other O2 molecules).

• In the tissues, haemoglobin binds to 2,3 DPG causing a

decrease in the affinity of Hb for O2, releasing oxygen to the
tissues.
• At the same time, CO2 binds to Hb due to high pressure of CO2 in
the tissues, reducing Hb affinity to oxygen (allosteric effect).

2,3 DGP enters between the two β chains and convert Hb from R
to T form, reducing the affinity of Hb for oxygen, this cause the
release of O2 (shift to the right on Hb dissociation curve)

*Note : HbF has no affinity to 2,3DGP, making the affinity of HbF

for oxygen more than HbA that has affinity for 2,3 DGP.
H+ (pH) and CO2 :
Formed from metabolic processes in tissues, bind to
Hb and decrease the affinity of Hb for O2 (allosteric
effect), this is also known as the Bohr effect.
https://www.youtube.com/watch?v=b2hKDxX-KjE