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
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:

Transports oxygen, CO
2
& protons
Its allosteric properties results from its
quaternary structures
A tetramer composed of pairs of different
polypeptides/subunits (o, |, , o 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
Hemoglobin function
Hemoglobin structure
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
o-helix (helix A to H)
Histidin F8 and E7 perform unique roles
in oxygen binding
Oxygen-binding curve for myoglobin is
hyperbolic

Heme
Heme Biosynthesis

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 5-
aminolevulinic acid. A series
of steps in the cytoplasm
produce coproporphrynogen
III, which re-enters the
mitochondrion. The final
enzymatic steps produce
heme.
Heme synthesis begins with condensation
of glycine & succinyl-CoA, with
decarboxylation, to form o-aminolevulinic
acid (ALA).

OOC CH
2
CH
2
C S-CoA
O
+

OOC CH
2
NH
3
+

OOC CH
2
CH
2
C
O
CH
2
NH
3
+
CO
2
CoA-SH
H
+ succinyl-CoA glycine
o-aminolevulinate (ALA)
o-Aminolevulinic
Acid Synthase
Synthesis of Porphobilinogen and Heme
http://themedicalbiochemistrypage.org/heme-porphyrin.html
Protoporphyrin IX
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
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):
c-
G
-
A
- |-o-|
Chromosome 16
(o-cluster):

2
-
1
-o
2
-o
1
-o
2
-o
1
-u
2
2 2 2 2
2
2
2
1
1 1
Globin Genes :
Hb types :
Embryo
(Gower-I) (Portland) (Gower-II)
Chains Synthesized
5'
3'
Chromosome #16
2 2
2 2
2 2
2 2
Fetus
Adult
(Hb-F)
(Hb-A) (Hb-A)
2
3'
5'
G
G
G
A
A
A
Globin Genes :
Hb types :
Chains Synthesized
Chromosome #11
50
30
10
6 18 30 6 18 30 42
prenatal age (wks)
% of total
globin
synthesis
birth
postnatal age (wks)
Types of hemoglobin
Hb Gower 1 = ,2c2
Hb Portland = ,22
Hb Gower 2 = o2c2
Hb Fetal (HbF) = o22
Hb Adult (HbA) = o2|2
Hb Adult minor (HbA2) = o2o2
Heme catabolism
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 CO
2
. 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).
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
Erythroid Class
X-linked sideroblastic
anemia, XLSA
-aminolevulinic acid
synthase 2, ALAS2
progressive iron
accumulation, fatal if not
treated
Congenital
erythropoietic
porphyria, CEP
uroporphyrinogen III
cosynthase
photosensitivity
Erythropoietic
protoporphyria, EPP
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 coproporphyrinogen oxidase
neurovisceral,
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
photosensitivity
Porphyria cutanea tarda type II, PCT
type II, also called the familial type
PCT, may also be referred to as
hepatoerythropoietic porphyria, HEP
uroporphyrinogen
decarboxylase in non-hepatic
tissues
photosensitivity
, some
neurovisceral