BIOSYNTHESIS &

CATABOLISM OF
HEMOGLOBIN
Abdul Salam M. Sofro
Faculty of Medicine
YARSI University Jakarta

Learning objectives
By the end of learning, students are
expected to understand:
Molecular structure and function of
hemoglobin
Biosynthesis of hemoglobin
Catabolic process and the fate of
hemoglobin catabolic products

Hemoglobin in blood

Blood cells development

General
Hemoglobin (four subunits) and its similar
molecule myoglobin (one subunit) are
iron-containing heme proteins consists
of apoprotein & non-protein heme
These heme proteins function in oxygen
binding, oxygen transport, electron
transport & photosynthesis carried
out by heme (a cyclic tetrapyrrole) & its
ferrous iron (at the center of the planar
ring)

Hemoglobin structure

Hemoglobin function

 The Molecular structure is similar

to Myoglobin :
MW 17,000 ; a monomer of protein with
153 AA residues
stores oxygen in red muscle tissue
will be released under condition of
oxygen deprivation (eg. Severe
aexercise) and used by muscle
mitochondria for ATP synthesis

 75% of the AA residues are present

in 8 -helix (helix A to H)
Histidin F8 and E7 perform unique
roles in oxygen binding
Oxygen-binding curve for myoglobin
is hyperbolic

 Hemoglobin:
Transports oxygen, CO2 & protons
Its allosteric properties results from its
quaternary structures
A tetramer composed of pairs of
different polypeptides/subunits (, , ,
etc. globin chains) a pair of globin chain
product of gene cluster in chromosome 11
& a pair of globin chain product of gene
cluster in chromosome 16

 Hb binds 2 protons for every 4 oxygen

molecules released & thus contributes
significantly to buffering capacity of blood
increase in proton concentration
promotes oxygen release, while increase
in PO2 promotes proton release.
At the lungs, oxygenation of Hb is
accompanied by expulsion and
subsequent expiration of CO2 Bohrs
effect (a reversible phenomenon with that
in the peripheral tissues)

 2,3-Bisphosphoglycerate (BPG) in Hb
Formed from glycolytic intermediate
1,3-bisphosphoglycerate
One molecule of BPG is bound per Hb
tetramer in the central cavity the
space is wide enough when Hb is in the
T form (deoxygenated)

 Binds more weakly to fetal Hb than to

adult Hb
Increase concentration of BPG lowers
the affinity of Hb for oxygen
(decreases P50) increasing the
ability of Hb to release oxygen at the
tissues

 As CO2 is absorbed in the blood, the

carbonic anhydrase (CA) in erythrocyte
catalyzes the formation of carbonic acid,
which in turn rapidly dissociate into
bicarbonate and a proton. To avoid
increasing the acidity of blood, a buffering
system must absorb these excess protons
this is carried out by Hb
CA

spontaneous

CO2 + H2O H2CO3 HCO3- + H+

 Mutant human Hb
Causes hemoglobinopathy (when biologic
function is altered)
Due to mutations in the gene that code
for globin chains:
Structurally abnormal Hb (HbM, HbS,
HbE, HbC etc)
Reduced synthesis of Hb
(thalassemias)
Diagnosed by special method (e.g.
molecular diagnosis)

Batak

Minahasa

1,5 0

Melayu
5,2 4,3

Minang

Bangka

3,7 2,9

5,4 4,5

Palembang
9,2 6,5

= trait hemoglobin-E

Palu

3,1 1,5

Banjar
0

1,2 3,7

1,2 6,1

2,9

Toraja
0

Tengger
0 10,6
Bali
Jawa
3,2 4,8

= trait thalassemia-

0 1,7

Dayak

4*

Sumbawa
5,1 6,8

Alor
0

Sasak
4,3 Sumba
2,5 36,6

Gambar . Pola distribusi dan prevalensi trait thalassemia- dan hemoglobin-E

pada berbagai populasi di Indonesia. * adalah hemoglobin OIna.

Heme

In addition to the heme b found in hemoglobin,

there are three different forms of heme found in
cytochromes such as those involved in the
process of oxidative phosphorylation.
Cytochromes of the c type contain a modified iron
protoporphyrin IX known as heme c. In heme c
the 2 vinyl (C=C) side chains are covalently
bonded to cysteine sulfhydryl residues of the
apoprotein. Only cytochromes of the c type
contain covalently bound heme. Heme a is also a
modified iron protoporphyrin IX. Heme a is found
in cytochromes of the a type and in the
chlorophyll of green plants

Biosynthesis of heme

Protoporphyrin IX

The sythesis of heme is a

complex process that involves
multiple enzymatic steps. The
process begins in the
mitochondrion with the
condensation of succinyl-CoA
and glycine to form 5aminolevulinic acid. A series
of steps in the cytoplasm
produce coproporphrynogen
III, which re-enters the
mitochondrion. The final
enzymatic steps produce
heme.

Synthesis of Porphobilinogen and

Heme

http://themedicalbiochemistrypage.org/heme-porphyrin.html

Globin
a polypeptide chain (protein)
Various types of polypeptide chain:

Alpha globin
Beta globin
Gamma globin
Delta globin
Epsilon globin
Zetta globin
Teta globin

Globin genes
Chromosome 11
(- cluster):
-G-A- --
Chromosome 16
(-cluster):
2-1-2-1-2-1-

C h ro m o s o m e # 1 6
5'

G lo b in G e n e s :
C h a in s S y n th e s iz e d
H b ty p e s :

2 2
(G o w e r- I)

2 2
(P o rtla n d )
E m b ryo

2 2
(G o w e r- II)

3'

C h ro m o s o m e # 11
5'

3'

G lo b in G e n e s :
C h a in s S y n th e s iz e d
H b ty p e s :

2 2
(H b - F )
F e tu s

2 2 2
(H b - A 2 ) (H b - A )
2

A d u lt

% o f to ta l
g lo b in
50
s y n th e s is

30

10
6

18

30

p re n a ta l a g e (w k s )

b ir th

18

30

p o s tn a ta l a g e (w k s )

42

Types of hemoglobin

Hb Gower 1
Hb Portland
Hb Gower 2
Hb Fetal (HbF)
Hb Adult (HbA)
Hb Adult minor (HbA2)

= 22
= 22
= 22
= 22
= 22
= 22

Catabolism of Heme

Heme breakdown
During its 120 day life span the erythrocyte
has traveled 200-300 miles. The process of
aging is called senescence.
Enzyme activity decreases (esp. glycolytic
enzyme which helps break down glucose,
the source of erythrocyte energy), and the
cell looses its deformability.

 MCHC (mean corpuscular hemoglobin

concentration) increases, the cell becomes
rounder, and the MCV mean corpuscular
volume) decreases.
90% of destruction of senescent Erythrocytes
occurs by extravascular hemolysis.
Macrophages of the mononuclear phagocyte
system remove them from circulation.

 Macrophages of the spleen are

especially active in removing aging, dead
and abnormal erythrocytes (e.g. cells
containing Heinz bodies or Howell-Jolly
bodies, siderocytes, target cells,
schistocytes, tear drop cells and
antibody-coated erythrocytes).

Normally, senescent red blood cells and

heme from other sources are engulfed by
cells of the reticuloendothelial system. The
globin is recycled or converted into amino
acids, which in turn are recycled or
catabolized as required.

Heme is oxidized, with the heme ring being

opened by the endoplasmic reticulum
enzyme, heme oxygenase. The oxidation
step requires heme as a substrate, and any
hemin (Fe3+) is reduced to heme (Fe2+)
prior to oxidation by heme oxygenase

Pathway for the

degradation of
heme to bilirubin.
Substituents:
M = methyl,
P = proprionic,
V = vinyl

 In individuals with abnormally high red

cell lysis, or liver damage with
obstruction of the bile duct, the bilirubin
and its precursors accumulate in the
circulation; the result is
hyperbilirubinemia, the cause of the
abnormal yellowish pigmentation of the
eyes and tissues known as jaundice.

 The protoporphyrin ring of heme is

disassembled and from the body. Its
alpha carbon is exhaled in the form of
CO2. The opened tetrapyrrole, biliverdin,
is converted to bilirubin which is then
carried to the liver by the plasma protein,
albumin.

 In the liver bilirubin is conjugated to

glucuronide to make it water soluble and
excreted along with bile into the intestines. In
the intestines it is converted by bacteria into
stercobilinogen and excreted in the stool;
some is eliminated as urobilinogen in the
urine.
Stercobilinogen and urobilinogen give feces
and urine their color.

 Unconjugated bilirubin (prehepatic) and

conjugated bilirubin (posthepatic) are
measured in serum as indirect
(unconjugated) and direct (conjugated)
bilirubin; used to monitor amount of
hemolysis.
Bilirubin and its catabolic products are
collectively known as the bile pigments.

Intravascular hemolysis
About 10% of normal erythrocyte
destruction occurs by intravascular
hemolysis.
In circulation the red cell is subjected to
metabolic and mechanical stresses:
turbulence, endothelial damage and fibrin
deposition, incompatibility due to
transfusion errors resulting in red cell
fragmentation (schistocytes) and/or
intravascular hemolysis.

 When the erythrocyte ruptures, hemoglobin

is released into the blood. The hemoglobin
dissociates into alpha-beta dimers and is
picked up haptoglobin, a protein carrier, to
prevent renal excretion of hemoglobin.
Haptoglobin carries the hemoglobin to the
liver for further catabolism where the
process proceeds as with extravascular
hemolysis.

 As haptoglobin is depleted, unbound

hemoglobin dimers appear in the plasma
(hemoglobinemia) and are reabsorbed by
the kidney up to a certain level and
converted to hemosiderin; beyond this level
hemoglobin shows up in the urine
(hemoglobinuria)
Intravascular hemolysis results in pink, red
or brown plasma (hemoglobinemia). Urine
may also show red color (hemoglobinuria).

http://diaglab.vet.cornell.edu/clinpath/modules/chem/images/bilirubin%20metabolism.jpg

Clinical Aspect of Heme

Metabolism
Clinical problems associated with heme
metabolism are of two types.
Disorders that arise from defects in the
enzymes of heme biosynthesis are termed
the porphyrias and cause elevations in the
serum and urine content of intermediates in
heme synthesis.
Inherited disorders in bilirubin metabolism
lead to hyperbilirubinemia

Porphyria

Enzyme Defect

Primary Symptom

-aminolevulinic acid
synthase 2, ALAS2

progressive iron
accumulation, fatal if not
treated

Erythroid Class
X-linked sideroblastic
anemia, XLSA
Congenital
erythropoietic
porphyria, CEP
Erythropoietic
protoporphyria, EPP

uroporphyrinogen III
cosynthase

photosensitivity

ferrochelatase

photosensitivity

Hepatic Class
ALA dehydratase deficient porphyria,
ADP

ALA dehydratase: also called

porphobilinogen synthase

neurovisceral

Acute intermittent porphyria, AIP

PBG deaminase: also called

hydroxymethylbilane
synthase or rarely
uroporphyrinogen I synthase

neurovisceral

Hereditary coproporphyria, HCP

neurovisceral,
coproporphyrinogen oxidase some
photosensitivity

Variegate porphyria, VP

protoporphyrinogen oxidase

neurovisceral,
some
photosensitivity

Porphyria cutanea tarda type I, PCT

type I, also called the sporadic type
PCT

hepatic uroporphyrinogen
decarboxylase

Porphyria cutanea tarda type II, PCT

type II, also called the familial type
PCT, may also be referred to as
hepatoerythropoietic porphyria, HEP

uroporphyrinogen
photosensitivity
decarboxylase in non-hepatic , some
tissues
neurovisceral

photosensitivity