CHAPTER

S3
in moo obn

Haemoglobin mol 6t000

dB-146 m ino
7
student should be able to
Competency: The
Specific Learning Objectives Describe synthesis and
functions of
I. Structure
1. (PY2.3)
haemoglobin
IL Some important definitions: Oxyhaemoglobin haemoglobin breakdown
2. (PY 2.3) Explain
carbamino-haemoglobin; reduced haemoglobin; Describe variants of haemoglobin
3. (PY 2.3)
carboxyhaemoglobin; methaemoglobin
III. Normal values
IV. Functions of haemoglobin
V. Disadvantages of 'free' haemoglobin
VI. Synthesis of haemoglobin Horizontal1
VII. Catabolism of haemoglobin integration:
VIIL Varieties of haemoglobin: HbA; Biochemistry
HbF-thalassaemia; HbS

in the RBCs is 3. Globin is a protein built from 4 polypeptide chains

The red, oxygen carrying pigment two 'a and two p' chains. Theretore, the normal
haemoglobin.It consists of the protein globin (polypeptide) adult haemoglobin (HbA) is written as HbA (.B)
united with the pigment haeni (heme).
Of two o-chains each contains 141 amino-acids and
of two B-chains each contains 146 amino-acids. Each
STRUCTURE chain is associated with one haem
polypeptide
1. Haem is an iron containing porphyrin, called group. Thus, there are
4 haem to the one molecule

iron-protoporphyrin X. The porphyrin nucleus isS of haemoglobin, contains 4 iron atoms and can carry

tetrapyrrole i.e. it consists of 4 pyrrole rings' joined 4 molecules (8 atoms) of oxygen.

The pyrrole
together by 4 methine (= CH-) bridges. 4. Oxygenation of 1st haem molecule in haemoglobin
and IV; the carbon atomms
rings arenumbered1, I, l and ô, the
increases the affinity of 2nd haem for oxygen and
of the methine bridges are labelled a, B, y Oxygenation of 2nd haem increases the affinity ot the
to which side chains are
position on pyrrole rings 3rd and s0 on. Therefore, the affinity of haemoglob
attached are numbered 1 to 8. The side chains at 1, 3, for the 4th oxygen molecule is many times that ot the
5 and 8 position are methyl (-CH,); 2 and 4 are vinyl
1st. This shifting affinity of haemoglobin for oxygen
(-CH CH,); 6 and
= 7 are propionic acid (-CH,CH,. results in:
COOH). (Fig. 7.1) () Sigmoid shape of oxygen-haemoglobin dissociation
The iron
2. The iron in haem is in the ferrous (Fe-") form. curve (page 430).
is attached to the 'N of each pyrrole ring. Each Fe
combines loosely and reversibly 2 3

with one molecule of oxygen. HC CH.CH, H,C CH.CH CH

0 Combination of haem with CH H

oxygen is called oXVgenation

and not oxidation, because,
NH
HC B CH Pyrrole Ring
after combination with oxygen,
iron in the haem stays in Fe
the oxygen does IV
state. Therefore, CH
but is H,C
not become 1onic oxygen CH, CH, COOH COOH.H,C.H,C
carried as molecular oxygen.

Fig. 7.1 Structure of haemoglobin molecule

58
 Ch. 7: Haemoglobin 59

Gi)Haemoglobin reacts with oxygen very rapidly

requiring less than 0.01 second. Similarly, (Fe)is oxidised to ferric (Fe3*) form and the cómpound
is called methaemoglobin. It is represented as HbOH.
deoxygenation of haemoglobin is also
5. Molecular weight of haemoglobin is 68,000.very rapid.
Disadvantages 1Sgm/d
) It cannot unite reversibly with gaseous oxygen.
SOME IMPORTANT DEFINITIONSS (Gi) It is dark coloured and when it is present in large
1. OXYHAEMOGLOBIN- quantities (more than 1.5 gm/dL) in circulation it
Haemoglobin reacts with
Oxygen to form resembles cyanosis i.e. blue colouration of skin.
oxyhaemoglobin
and is represented as
HbO, Some oxidation of haemoglobin to methaemoglobin
occurs normally, but an enzyme in RBCs, the
affinity of haemoglobin for oxygen is influenced
The
by pH, temperature and concentration of 2,3, diphospho- NADH (dihydronicotinamide adenine dinucleotide)-
glycerate (2,3 DPG) in the RBCs, a product of metabolism methaemoglobin reductase system, converts methaemo-
of glucose. As globin back to haemoglobin. Congenital absence of this
concentration of 2,3 DPG rises, the affinity system causes hereditary methaemoglobinemia, a fatal
of haemoglobin for oxygen falls and the
oxygen-haemoglobin
dissociation curve is shifted to the right; as a result more condition. (Fig. 7.2)
NADU
Oxygen is released by blood to the tissues. Saviov
IMPORTANT NOTES
1. At high altitude (eg. 5000 metres above sea level)
2,3 DPG concentration in RBCs increases by 50%
and this makes more oxygen available to the
tissues.
2. Stored blood loses its 2,3 DPG and oxygen
affinity
of haemoglobin increases resulting in less release of
ig. 7.2 Methaemoglobinemia
Oxygen.
(Note: Dark coloured fingers on the right)
2. CARBAMINO-HAEMOGLOBIN NORMAL VALUES
Carbon dioxide reacts with haemoglobin to form carbamin0- 1. At birth: 23 gm/dL, because RBC count is more.
2. At the end of 3 months:
haemoglobin. 10.5 gm/dL,
as an infant is
totally on milk feed which is devoid of iron.
CO,+ HbNH, HbNH COOH 3. After 3 months,
haemoglobin increases gradually and
at the end of 1 year it
3. REDUCED (DEOXYGENATED) 4. Adults
becomes 12.5 gm/dL.
HAEMOGLOBIN Males 14-18 gm/dL (Average: 15.5 gm/dL)
Haemoglobin from which oxygen has been removed Females 12-15.5 gm/dL (Average:
is called reduced or deoxygenated haemoglobin and is 14 gm/dL)
represented as Hb. Clinically, 14.8 gm/dL haemoglobin irrespective of
sex is
regarded 100% haemoglobin.
as

4. CARBQXY HAEMOGLOBIN When blood is equilibrated with 100%

oxygen
(pO, 100 mmHg), the normal haemoglobin becomes
=
or CARBON MONOXY HAEMoGLOBIN
100% saturated.
Carbon monoxide (CO) reacts with haemoglobin to
form carboxy haenmoglobin or carbon monoxy haenmoglobin.
1
gm/dLhaemoglobin when fully saturated combines
with 1.34 mL oxygen, therefore,
The affinity of haemoglobin for CO is 210 times than its haemoglobin
concentration is an index of oxygen-carrying
affinity for oxygen which consequentlydisplaces oxygen of blood. capacity
on haemogobin, reducing the oxygen carrying capacity 1.34 m oo
of blood. Normal values -

males: 21 mL/dL, females: 18

mL/dL.
5. METHAEMOGLOBIN
FUNCTIONS OF HAEMOGLOBIN
When either reduced_or oxygenated haemoglobin is 1. Facilitates transport of
oXVgen from lungs to the
agents, the ferrous tissues.
exposed to various drugs or oxidising
 V1
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Unit I1: Blood
60 adequate
amount occu
is required;
copper contain tra
the tissues to the iron preparations
2. Facilitates transport of CO, from diet and
most

of copper.
lungs. tor manufacture of
acid-base buffer, being protein.
a necessary
is
3. Itacts as an excellent (1l) Cobalt
in_the umen
of whole bacterial action
It is responsible for 70% buffering power vitamin B by increases the production of
also
blood. of GTL. It which, in turn, stimulate
flow and BP
4. It plays role in regulation of blood
a hormone erythropoietii
nitric oxide (NO) binding of RBCs.
Haemoglobin has additional the development
from GT
increased by O. Therefore, increases iron absorption
site on the P-chain which is releases (iv) Calcium
-

NO in the lungs and C, vitamin B, and foli

haemoglobin binds with vasodilation. 3. Role of vitamins Vitamin
-

acid which in tu
nucleic
it in the tissues where it promotes of
acid help in synthesis of RBCs, Vitamin C
is required for
the development
DISADVANTAGES OF FREE of iron by reducing ferric (Fe
also helps absorption
HAEMOGLOBIN (Fe-") compounds.
to forms
is contained within the RBCs?)
(Why haemoglobin
dissolved in the plasma (called
1. If haemoglobin was CATABOLISM OF HAEMOGLOBIN
would lead to:
free haemoglobin) it 'tissue-macrophage system
plasma, hence of destroyed in
1) increase in the viscosity of Old RBCs are

whole blood, causing BP to

rise, and (page 114)
to destruction shown
increase in the osmotic' pressure of plasma The fate of haemoglobin after its
is
(11)
100 mmHg in Fig. 7.3.
with the mechanism of
) and (i) interfere
and tissue
fluid exchange between capillaries NOTE
spaces in urine Approx. 0.3gm of haemoglobin
is destroyed and
2 Loss of free haemoglobin_ hy the kidneys 0.3gm synthesized every hour.
(haemoglobinuria)also results in kidney damage.
3. Free haemoglobin is taken up and rapidly destroyed
by the tissue-macrophage system.
Haemoglobin
SYNTHESIS OF HAEMOGLOBIN
Tissue-macrophage system
Synthesis of haemoglobin requires the provision of nutrients (split offinto)
eg proteins, vitamins, minerals (specially iron). It only
takes place in the developing RBCs (erythroid series).
GLOBIN HAEM
RE-USED
(enters amino- iron protoporphyrin IX)
Factors controlling haemoglobin formation for
acid pool)
1. Role of proteins A low protein intake decreases synthesis
(split off of
haemoglobin regeneration even in the presence of
excess of iron; the limiting factor hereislack of globin
formation. Remaining part Fe2
2 Role of Minerals
) Iron (Oxidised combines with
(a) it helps in formation of lhaem;
by haem Apoferitin
otygenase) (Tissue protein)
(b) iron content of haemoglobin is 0.33%, therefore
100 mL of blood containing 15 gm of BILIVERDIN FERRITIN
haemoglobin contains 15x 0.33/100 = agreen pigment (stored in the
approx. 50 mg of iron. (Tetrapyrrole straight chain liver)
free from
(c) as life span of RBCis120days, therefore, 0.8% globin and iron)
(1/120x 100) of total blood haemoglobin reduced by
contained in 50 mL (6 litres x 0.8%) of blood, biliverdin reductase
is destroyed daily, releasing approx. BILIRUBIN
25 mg of a yellow/orange pigment
iron. This iron is reused for fresh
synthesis (Excreted in bile)
ofhaemoglobin.
(i) Copper helps in promoting the absorption,
mobilization and utilization of iron. Very little
Fig 7.3 Fate of
haemoglobin in the body
 Ch.7: Haemoglobin 61

VARIETIES OF HAEMOGLOBIN (i) It is much more resistant to the action of alkalies

than

Several varieties of haemoglobin occur in human, in all HbA. This property is made use of in a photoelectric
the haem moiety 1s the same; physical and chemical colorimetric method to estimate HbF in the presence
differences being due to variations in the composition of HbA.
of the peptides of the 'globin fraction. The amino-acid (1) HbF has greater affinity for oxygen, because of poor
sequence in the polypeptide chains of haemoglobin is
binding of 2,3-DPG to the Ypolypeptide chain,
determined by 'globin genes. therefore, it can take much larger volume of oxygen
than HbA at low oxygen pressure. This facilitates
IMPORTANT NOTE HbPequcb the movement of oxygen from maternal to foetal
circulation. HbF is 70% saturated at 20 mmHg of
In diabetes mellitus (DM) patients, small amount
of HbA is non-enzymatically glycosylated to form pO2 pressure (pO,), whereas HbA is only 30-35%
saturated at this pressure.
haemoglobin Ajc (HBA). It has agucose attached
iv) Its life span is less (about 80 days) as compared to
to the terminal valine in each B-chain.
HbA1c that of HbA (120 days).
concentrations are measured as an index of control of
(v) At birth HbE predominates (approx. 80o), it
DM (page 660).
Can be ustd lo gradually disappears 2-3 months after birth.
mess
Hence, persistence of HbF beyond the age of 4-6
1. ADULT HAEMOGLOBIN (HbA) months after birth should raise the suspicion of disordered
It is of two types: MbA synthesis of HbA due to deficient production of a or
) Haemoglobin A (oB,) - predominately seen; as
B-chains (called oa or B Thalassaemia respectively). The
condition is associated with severe anaemia in children
described above. and require frequent blood transfusion.
i) Haemoglobin A, (a,8,). Here B chains are replaced
by o chains. The ô chains also contain 146 amino-acid
but 10 individual amino-acids diliier from those in
Thalassaemia
the B chain. Approx. 2.5% of the total haemoglobin is
HbA, in normal adults. It produces no abnormality B (more common) a (rare)

and is regarded as normal haemoglobin.

HbA appears in foetus, after 5 months (20 weeks) of Major Minor
intra-uterine life, when bone marrow begins to function (more common)
as a haemopoietic agent.
The main differentiating features between the
Amount of HbA, of the major
and minor B-thalassaemia are give in Table 7.1.
Age total haemoglobin

20 weeks of intra-uterine 6% (rest is HbF) 3. HAEMOGLOBIN-S (Hbs)

At
life It is inherited as Mendelian dominant.
Here, in each
B-polypeptide chain of HbA at position 6, one glutamic acid
At birth 20% is replaced by a valine. Therefore, when HbS is reduced
(eg.
as in low O, tension or low
At 2 months (post-natal) 50% pH at tissue levels), it becomes
much less soluble than HbA, haemoglobin
into crystals within RBCs causes the
precipitates
90%
At 4 months
(post-natal) following:
(i) It damages cell membrane producing increased
More than 1 year 99% (< 1% is HbF)
fragility of RBCs.

2. FOETAL HAEMOGLOBIN (HbF - a2Y2)

(ii) Crystals elongate and RBCs become sickle
shaped
(reaping hook shape - Fig. 8.10, page 74) which decreases

the blood flow to tissues due to

Salient features phenomenon of
sickling (increased blood viscosity).
(1) Its structure is same as of HbA, except that the
P-chains are replaced by y-chains. -chains
also () and (i) cause RBCs to become more fragile producing
contain 146 amino-acids but have 37
amino-acids severe anaemia, called sickle cell anaemia. Finally, patient
that differ from those in the B-chains. dies in a few days due to severe anaemia and
secondary
infection.
Wb Mbous Wbso.
B3