HAEMOGLOBIN: BIOCHEMISTRY AND

GENETICS
DR. CROSDALE O. PUGHIKUMO
MBBS, PGDM, FMCPath (Haem), MSc.

COLLEGE OF HEALTH SCIENCES

NIGER DELTA UNIVERSITY

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 Introduction
• Haemoglobin (Hb) is the red pigment contained in the red cells.
• It is a complex molecule made up of protein and iron with a MW of
68,000 D.
• It constitutes about a third of the red cell mass.
• Its oxygen-binding capacity is about 1.34 mL O2/g.
• This satisfies the average adult consumption of about 250 mL/min.

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 Structure of Haemoglobin
• Hb has a heme and a globin component.
• Hb has 4 polypeptide chains joined together by disulphide bridges.
• Located at the centre of the chain is a heme group which contains
iron and it is capable of binding one molecule of oxygen.
• Ferrous iron is attached to a Nitrogen atom in each of the Pyrrole
rings.
• The globin chain has 4 sub-units. In the normal adult Hb (Hb A), this is
constituted by two α chains of 141 amino acids each and two β
chains of 146 amino acids each.

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• The polypeptide chains are coiled up with hydrophilic R groups
outwards interacting with H2O via hydrogen bonding and hydrophobic
ends inwards making it soluble.
• 4 haem units are attached to one globin unit.
• Thus, one Hb molecule contains 4 iron atoms in Fe2+ form which is
capable of binding 4 oxygen molecules.

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 Synthesis of Haemoglobin
• Haem synthesis occurs in virtually all cells but most importantly in the
liver (for cytochromes), muscles (for myoglobin), and red cell
precursors.
• The synthesis of the haem molecule and the globin chains occurs at
two different sites in the cell but in an organised manner.
• The Globin chains are produced in the ribosomes.
• Haem is produced in the mitochondria.

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 Steps in haem synthesis
• 1.Glycine and succinate condense to form 5-aminolaevulinate in the presence
of Pyridoxal phosphate. This step is energy-dependent and rate-limiting
catalysed by ALA synthase.
• 2. Condensation of 2 molecules of ALA forms Porphobilinogen (PBG) a
monopyrrole. This and other reactions occur in the cytoplasm.
• 3. Four of PBG then forms Uroporphyrinogen, a tetrapyrrole with 2 isomers I
and III. III is the major pathway.
• 4. The following are then formed in succession: Corproporphyrinogen III which
enters the mitochondria forming Protoporphyrinogen IX and Protoporphyrin
IX.
• Ferrous iron, Fe2+ is added to Protoporphyrin IX to form Haem.

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 Disorders of haem synthesis: The Porphyrias

• They are a rare group of inherited disorders of haem synthesis arising

from a deficiency of enzymes in the synthetic pathway.
• Haem production is impaired.
• The major clinical features are:
• Neurologic disturbances like acute abdomen and peripheral
neuropathy occurs when ALA and PBG accumulates.
• Cutaneous manifestations: like photosensitivity, blisters in exposed
parts when sunlight activates excess Porphyrins.

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 Classification of Porphyrias

• This is based on the main site of accumulation of Porphyrins:

• 1. Hepatic porphyrias:
• A. Acute porphyrias: Acute intermittent porphyria
Hereditary coproporphyria
Porphyria variegate
• B. Chronic porphyrias: porphyria cutanea tarda: genetic or acquired.
• 2. Erythropoietic porphyrias: congenital erythropoietic porphyria
Protoporphyria.

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 Globin synthesis

• The globins are single chain polypeptides produced under genetic

control like all other proteins.
• The genes encoding the globin chains are present in 2 clusters:
• The α cluster is located in the short arm of chromosome 16. It has
two types; the α-chain and the ζ-chain genes.
• The genes for the β cluster are located in the short arm of
chromosome 11. There are 4 types; β, γ, δ and ε genes.

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 Succession of globin chains

• Production of globin chains commences as early as the third week of

gestation.
• A. Embryonic chains: a. Haemoglobin Gower 1:(ζ2ε2)
• b. Haemoglobin Portland: (ζ2γ2)
• c. Haemoglobin Gower 2: (γ2ε2)
• B. Foetal chains: Haemoglobin F (α2γ2)
• Haemoglobin A (α2β2)
• C. Adult chains: HbF, HbA, HbA2 (α2δ2).
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 Normal hemoglobins

Embryonic Fetal Adult

Hemoglobi hemoglobi hemoglobi
ns n ns
●
Hemoglobin A- alpha
●
Gower 1- zeta (2), (2), beta (2) ~95%~
epsilon (2)
●
~
●
Hemoglobin F- ●
Hemoglobin A2-
●
Gower 2- alpha
alpha(2), gamma alpha(2), delta(2)
(2), epsilon (2) ~1.5-3.7%
(2) ●
●
Portland-Zeta (2), ●
Hemoglobin F-
gamma (2) alpha(2), gamma (2)
<2 %

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Hemoglobins in normal adults

a b a g a d

b a g a d a

Hb A
HbA98%
Hb F
1% HbA
Hb A2
3.5% 2

98% ~1% <3.5%

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 Major functions of haemoglobin

• 1. Transport of oxygen from the lungs to the tissues.

• 2. Transport of CO2 from the tissues to the lungs.
• 3. Maintenance of acid-base balance.

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 Haemoglobin-Oxygen dissociation curve

• Oxygen binds to haemoglobin reversibly via covalent bonding.

• When one molecule of oxygen binds heme, a conformationa change
occurs, the β-chains move closer and 2,3-DPG is expelled.
• This increases the affinity of heme for oxygen making it easier for the
2nd, 3rd and 4th molecules of oxygen to bind.

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 The Hb-O2 dissociation curve

• The process is reversed when oxygen is released to the tissues.

• The oxygen-carrying capacity of Hb varies with PO2.
• The PO2 at which Hb is half-saturated is 26.6mmHg.
• This results in the sigmoid shape of the Hb-O2 dissociation curve.

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 Factors affecting the position of the curve

• 1. Structure of haemoglobin.
• 2. pH
• 3. Concentration of CO2 in the red cells.
• 4. Concentration of 2,3 DPG in the red cells.

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 Causes of a right shift

• 1. Haemoglobin S
• 2. High CO2
• 3. High H+
• 4. High 2,3-DPG.
• 5. High temperature.

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 Causes of a left shift

• 1. Low 2,3 DPG levels.

• 2. Low CO2
• 3. Low H+
• 4. Haemoglobin F.
• 5. Low temperature.

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 Hb transport of CO2: The Bohr effect

• The Bohr effect explains the major gas exchange mechanism in the
body.
• Hb’s O2-binding affinity is inversely related both to acidity and CO2
concentration.
• The release of oxygen at the level of the capillaries in the tissues
facilitates binding of CO2 to haemoglobin.
• In the lungs, the release of CO2 facilitates the binding of O2 to
haemoglobin.
• About 23% of CO2 is transported this way.
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 Bohr effect contd.

• Alterations in blood pH, shifts oxygen dissociation curve

• In acidic pH, the curve shifts to the right
• Results in an enhanced capacity to release O2 where it is needed

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Nonfunctional hemoglobins

• Carboxyhemoglobin
• Oxygen molecules bound to heme are replaced by carbon monoxide.
• Slightly increased levels of carboxyhemoglobin are present in heavy smokers and as a result of environmental
pollution.
• Can revert to oxyhemoglobin.
• Methemoglobin
• Iron in the hemoglobin molecule is in the ferric (Fe3) state instead of the ferrous (Fe2) state.
• Incapable of combining with oxygen.
• Can occur as a result of strong oxidative drugs or to an enzyme deficiency.
• Can revert to oxyhemoglobin
• Sulfhemoglobin
• Hemoglobin molecule contains sulfur.
• Caused by certain sulfur-containing drugs or chronic constipation.
• Cannot revert to oxyhemoglobin and may cause death.

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HAVE A BLESSED DAY

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