Haemoglobin

Haemoglobin is a
conjugated protein present
inside RBC. It is composed
of haeme, an iron
containing compound
belonging to
protoporphyrin and globin
belonging to class of
protein called globulins.
 Functions of Hemoglobin

It is required for :
1. Transport of oxygen : oxygen binds with
haemoglobin to form oxyhaemoglobin which
is an unstable compound. This combination is
reversible and oxygen can be released from
the compound to form reduced haemoglobin.
2. Transport of carbon dioxide : haemoglobin
binds with carbon dioxide to form
carboxyhaemoglobin which is reversible and
carbon dioxide can be released from it.
Affinity of haemoglobin for carbon dioxide is
twenty times more than that for oxygen.

3. As a blood buffer
 Derivatives of Haemoglobin

Oxyhaemoglobin :
It is a compound of haemoglobin with oxygen .
Iron remains in the ferrous (Fe++) state in
oxyhaemoglobin. It is not a stable compound .
Methaemoglobin :

It is also a compound of Hb with Oxygen . It is

a stable compound . Oxygen cannot be
removed. Iron remains in the ferric state .

Carbohaemoglobin :

It is a compound of Hb with CO2 , by union of

CO2 the globin portion.
Carboxyhemoglobin or Carbomonoxy haemoglobin :

Haemoglobin combined with CO instead of oxygen .

It is present in blood in coal gas poisoning.

Sulphaemoglobin :

It is formed by the combination of Hb with H2S . The

compound is very stable and is sometimes found in
the blood after certain kinds of drug poisoning.
Normal concentration
• Males - 14 - 17 g /100 ml

• Females - 12 - 16 g /100 ml
 Structure of Haemoglobin

Haemoglobin is a conjugated protein. It consists of a

protein part, globin and an iron containing
pigment, haeme. Iron is present in unstable ferrous
form. Pigment part is porphyrin formed by four
pyrrole molecules attached to one another by
methine bridges (tetrapyrrole ring). Globin contains
four polypeptide chains, two a and two b chains.
 Formation of Haemoglobin

Synthesised by cells of erythroid series in bone

marrow. Hemoglobin appears in the
intermediate normoblast stage of the red blood
cells. Succinyl - CoA combines with glycine
eventually to form a pyrrole molecule.
Four pyrrole molecules combine to form
protoporphyrin IX, which then incorporates
iron to form the haeme molecule. Each haeme
molecule combines with a long polypeptide
chain of globin to form the hemoglobin chain
and four such chains form a haemoglobin
molecule.
The most common form of haemoglobin in the adult human
being, haemoglobin A, is a combination of two alpha chains
and two beta chains. Haemoglobin A has a molecular
weight of 64,458. In each haemoglobin molecule there are
four iron atoms, each of which can bind loosely with one
molecule of oxygen, making a total of four molecules of
oxygen (or eight oxygen atoms) that can be transported by
each haemoglobin molecule. The types of haemoglobin
chains in the haemoglobin molecule determine the binding
affinity of the haemoglobin for oxygen.
 Combination of Haemoglobin with
Oxygen.
The most important feature of the hemoglobin
molecule is its ability to combine loosely and
reversibly with oxygen. The primary function of
hemoglobin in the body is to combine with oxygen
in the lungs and then to release this oxygen readily
in the peripheral tissue capillaries, where the
gaseous tension is lower than in the lungs.
In methaemoglobinaemia, iron in

haemoglobin is oxidised to ferric state and

oxygen can not release itself from

haemoglobin. Haemoglobin combines with

carbon dioxide to serve as a blood buffer.

 Iron Metabolism

Iron is important for the formation not only of

hemoglobin but also of other essential elements in
the body (e.g., myoglobin, cytochromes, cytochrome
oxidase, peroxidase, catalase). The total quantity of
iron in the body averages 4 to 5 grams, about 65
per cent of which is in the form of hemoglobin.
Absorption, Transport and Storage of
Iron.
Iron is absorbed from all parts of the small
intestine, especially upper part including
duodenum. The liver secretes apotransferrin (a
beta globulin) into the bile, which binds with
free iron to form transferrin.
It is absorbed into the epithelial cells and later
released into the blood capillaries in the form
of plasma transferrin. Iron absorption from
the intestines is extremely slow, at a
maximum rate of only a few milligrams per
day.
The iron is loosely bound in the transferrin and
can be released to any tissue. A unique
characteristic of the transferrin molecule is
that it binds strongly with receptors in the cell
membranes of erythroblasts in the bone
marrow and delivers the iron directly to the
mitochondria, where heme is synthesized.
Excess iron in the blood is deposited especially in
the liver hepatocytes and less in the
reticuloendothelial cells of the bone marrow. In
the cell cytoplasm, iron combines mainly with a
protein, apoferritin, to form ferritin. This iron
stored as ferritin is called storage iron and small
quantity is stored as hemosiderin.
 Fate of Haemoglobin

After a life span of 120 days RBC burst and

release the hemoglobin which is phagocytized by
macrophages, especially by the Kupffer cells of
the liver and macrophages of the spleen and
bone marrow. Within the phagocytes, haeme is
converted into a straight chain compound and
release iron from the hemoglobin into the blood.
The tetrapyrrole straight chain compound thus
formed is called biliverdin.Biliverdin is
oxidized to form bilirubin. Bilirubin now
comes out of the phagocyte and combine with
albumin to be transported in the plasma as
bilirubin - albumin complex. This complex is
called free bilirubin.
The free bilirubin ultimately enters the liver and
here the albumin is removed and the bilirubin is
conjugated with glucoronic acid to form
bilirubin glucoronide, which is transported to
duodenum through biliary canaliculi. Here it is
unconjugated and reduced to form urobilinogen
and stercobilinogen which are excreted via urine
and stool.
 Daily Loss of Iron
A man excretes about 0.6 milligram of iron each day, mainly
into the feces. Additional quantities of iron are lost when
bleeding occurs. For a woman, additional menstrual loss of
blood brings long term iron loss to an average of about 1.3
mg/day. Conversely, when the iron stores have become
depleted, the rate of absorption can accelerate probably five
or more times normal. Thus, total body iron is regulated
mainly by altering the rate of absorption.
 Hemoglobinopathies
The hemoglobinopathies are a group of
conditions in which the structure or production
of haemoglobin has been altered in some way.
Haemoglobin in normal individuals are mainly
HbA1, HbA2 and HbF. During foetal life HbF is
main haemoglobin which is replaced by HbA
within first six months of life
HbA1 contains two a and two b chains, HbA2
contains two a and two d chains and HbF
contains two a and two g chains. When the
sequential order of amino acid is altered or
the chain is altered by addition or deletion,
abnormal haemoglobin is formed.

Eg : HbS, HbC, HbE, HbM etc.

Sickle cell syndrome :

Sickle -cell haemoglobin

(HbS) is the major
abnormal haemoglobin in
this group. The 6th amino
acid of the beta chain is
valine instead of the normal
glutamic acid of HbA.
When this hemoglobin is exposed to low
concentrations of oxygen, it precipitates into
long crystals which elongate the cell giving
the appearance of a sickle. It also damages the
cell membrane, so that the cells become
highly fragile, leading to serious anemia
Such patients frequently experience a vicious
circle of events called a sickle cell disease
‘crisis’, in which sickling leads to rupture of
red cells, which causes a further decrease in
oxygen tension and still more sickling and red
cell destruction. Symptoms occur only after 6
months of age.
Early symptoms are anaemia, hepatosplenomegaly,
growth retardation, recurrent infections and painful
swelling of hands. Sickling crises are characterised by
painful episodes associated with constitutional
disturbances like fever, worsening of anaemia and
other organ symptoms. Cardiomegaly, cardiac
murmurs, pulmonary hypertension and cor
pulmonale are late manifestations.
 Defective haemoglobin synthesis

Thalassemias :

Genetic disorders in which synthesis of normal

polypeptide chains forming adult haemoglobin are
suppressed. Synthesis of either the a chain or b chain is
not sufficient in amount. Thalassemia can be major or
minor.
In b thalassemia major ( Cooley’s anaemia ),
synthesis of b chain is insufficient. They will
have relative excess of a chains and as
production of g chain continues, foetal
haemoglobins are formed. Child is normal at
birth and for few months.
Anaemia appears gradually from 6 - 24th
week. Abdominal protrusion from gross
hepatosplenomegaly, thin limbs and the
classic thalassemia facies. Haemoglobin is
generally 3 - 4 g/dl. erythrocytes are
hypochromic and microcytic.
Anemias
Anemia means deficiency of
hemoglobin in the blood, which can
be caused by either too few red
blood cells or too little hemoglobin
in the cells.
Blood Loss Anemia :
In rapid hemorrhage, the replacement of fluid
portion of the plasma, lowers concentration of red
blood cells. In chronic blood loss, the red cells
produced are much smaller and have too little
hemoglobin as absorption of iron from the
intestines is not as rapid as its loss .This results in
microcytic, hypochromic anemia.
Iron Deficiency Anaemia :

Major cause of iron deficiency is chronic

blood loss; in addition iron deficient diet
or lack of absorption of iron contribute
to it. Women of reproducing age and
children are susceptible to iron
deficiency. RBC count may be near
normal, but Hb %, PCV, MCV values are
low. Anemia is microcytic hypochromic.
Aplastic Anemia :

Bone marrow aplasia means lack of functioning

bone marrow. Exposure to gamma ray radiation
causing complete destruction of bone marrow
result in lethal anemia. Excessive X-ray
treatment, certain industrial chemicals, and even
drugs to which the person might be sensitive can
cause the same effect.
Megaloblastic Anemia :

Deficiency of vitamin B12, folic acid, and

intrinsic factor from the stomach mucosa etc.
can lead to slow reproduction of erythroblasts
in the bone marrow. As a result, the red cells
grow too large, with odd shapes, and are called
megaloblasts.
Atrophy of the stomach mucosa, as in
pernicious anemia or surgical total
gastrectomy can lead to megaloblastic anemia.
In such conditions the erythroblasts formed
are mostly oversized with bizarre shapes and
fragile membranes. These cells rupture easily.
Hemolytic Anemia :

Different abnormalities of the red blood cells,

many of which are hereditarily acquired,
make the cells fragile and rupture easily as
they go through the capillaries, especially
through the spleen.
In hereditary spherocytosis, the red cells are
very small and spherical and as they do not
have the loose, bag like cell membrane
structure of the normal biconcave disc they
are easily ruptured by even slight
compression on passing through the splenic
pulp and some other tight vascular beds.
 In erythroblastosis fetalis, Rh-positive red
blood cells in the fetus are attacked by
antibodies from an Rh-negative mother. These
antibodies make the Rh-positive cells fragile,
leading to rapid rupture and causing the child
to be born with serious anemia.

RBC count, PCV low. MCV low or normal.

 Signs and Symptoms

General symptoms like fatigue, disinclination to

work, mental apathy, pallor, exertional dyspnoea,
effort angina and cardiac failure may occur due to
diminished oxygen carrying capacity of blood and
hyperdynamic circulation. Alimentary symptoms
are loss of appetite, constipation and abdominal
distention. Liver may be enlarged and tender.
Cardiovascular changes include tachycardia,
cardiomegaly, high volume pulse, prominent
third heart sound, ejection systolic murmurs
and signs of cardiac failure. Pallor is marked
over mucous membranes, skin and nails.
Neurological manifestations include apathy,
loss of mental alertness, paraesthesia over
extremities and brisk tendon reflexes.
 Polycythemia Vera
Polycythemia vera is caused by a genetic
aberration in the hemocytoblastic cells that
produce the blood cells. Increasing blood viscosity
in polycythemia, causes very sluggish blood flow
through the peripheral blood vessels, a larger than
normal quantity of hemoglobin is deoxygenated.
Therefore, a person with polycythemia vera
ordinarily has a ruddy complexion with a bluish
(cyanotic) tint to the skin.