Hemoglobin Metabolism

Learning Objectives

Describe biosynthesis of heme, mention its functions in

the body and disorders of heme biosynthesis

Describe heme catabolism

Describe disorders of bilirubin metabolism

Describe laboratory tests done to differentiate jaundice.

 Heme is an iron containing porphyrin.
 Heme is the prosthetic group of several proteins and
enzymes including hemoglobin, myoglobin,
cytochrome, cytochrome P450, enzymes like catalase,
certain peroxidase, and tryptophan pyrrolase.
 Heme is synthesized from porphyrin and iron
 Porphyrin ring is coordinated with an atom of iron to
form heme
Figure 14.1: Heme molecule showing a pyrrole molecule and an
atom of iron (Fe 2+) in the center of the porphyrin ring,
which is tetrapyrrole structure.
(M: methyl; P: propionyl; V: vinyl)
 Biosynthesis Of Heme
 Heme synthesis takes place in all cells (except in mature
erythrocytes), but occurs to the greatest extent in the
bone marrow and liver.
 Biosynthesis of heme may be divided into three stages:
 Stages of Heme Synthesis

1. Biosynthesis of δ-aminolevulinic acid (ALA) from

precursor glycine and succinyl-CoA

2. Formation of porphobilinogen (PBG) from δ-
aminolevulinic acid
3. Formation of porphyrins and heme from
porphobilinogen.
Regulation of Heme Synthesis

 δ-aminolevulinic acid synthase (ALAS), a

mitochondrial allosteric enzyme that catalyzes first

step of heme biosynthetic pathway, is a regulatory

enzyme. It is feedback inhibited by heme.

 Regulation also occurs at the level of enzyme synthesis.

• Increased level of heme represses the synthesis of

δ-aminolevulinic acid synthase

• Decreased level of heme induces the synthesis of δ-

aminolevulinic acid synthase.

 The iron atom itself activates heme synthesis.

 Several drugs induce the synthesis of hepatic ALAS,

e.g.steroid hormone métabolites, éthanol, barbiturates,

Disorders of heme biosynthesis

Disorders of heme biosynthesis may be genetic or

acquired.

1. The most common acquired defect is lead poisoning.

Lead can inactivate d-ALA dehydratase and ferrochelatase

by combing with essential thiol groups of these enzymes .

Signs of lead poisoning include
 Elevated levels of protoporphyrin in erythrocytes
 Elevated urinary levels of ALA and coproporphyrin III.
 Production of heme is decreased.
 Anemia results from lack of hemoglobin and
 Energy production decreases due to lack of cytochromes
for the electron transport chain.
2. X-linked sideroblastic anemia (XLSA):
 It is a congenital disorder due to a defect in δ-
aminolevulinate synthase (ALAS), which is involved
in the first step of heme synthesis.
 Sideroblastic anemia is a form of anemia in which the
bone marrow produces ringed sideroblasts rather than
healthy red blood cells.
 Sideroblasts are nucleated erythroblasts (precursors to
mature red blood cells) with granules of iron
accumulated in the mitochondria surrounding the
nucleus.
 In sideroblastic anemia, iron is available but cannot
incorporate into hemoglobin.
 Symptoms of sideroblastic anemia include skin
paleness, fatigue, dizziness, and enlarged spleen and
liver.
3. Porphyrias are a group of genetic diseases that result
from defect in enzymes of the biosynthetic pathway
from glycine to porphyrins. In porphyrias, specific
porphyrin precursors accumulate in erythrocytes,
body fluids, and in the liver.
 The Porphyrias
 Porphyria is named from the ancient Greek word
porphura, meaning purple. Purple color is caused by
pigment-like porphyrins in the urine of some patients with
defects in heme synthesis.
 The porphyrias are rare group of inherited disorders
resulting from deficiencies of enzymes in the biosynthetic
pathway for heme, and lead to accumulation and
increased excretion of porphyrins or porphyrin precursors
(ALA and PBG).
 With the exception of two, congenital erythropoietic
porphyria and aminolevulinic acid dehydratase
deficiency porphyria (ALAD Porphyria), which are
genetically autosomal recessive diseases, all the
porphyrias are inherited as autosomal dominant
disorders.
 Aminolevulinic acid dehydratase deficiency porphyria
(ALAD Porphyria) is a hepatic and extremely rare
congenital condition.
 Most individuals inheriting this condition are
heterozygotes and are usually asymptomatic, because
the single copy of the normal gene provides a sufficient
level of enzyme function.
 However, certain nutritional and environmental factors
can cause a formation of δ-aminolevulinate and
porphobilinogen, leading to attacks of acute
abdominal pain and neurological dysfunction.
Classification of porphyria

The hereditary porphyrias are classified into two categories

on the basis of the organs or cell that are mostly affected.

Thus, porphyria are classified into:

1. Erythropoietic porphyria : Enzyme deficiency occur in

erythropoietic cells of the bone marrow.

2. Hepatic porphyria : Enzyme deficiency occur in

hepatic cell.
Figure 14.3: Porphyrias caused by mutations
of enzymes.
 Biochemical Basis of the Major Signs and Symptoms of
Porphyrias
 Where the enzyme lesion occurs early in the pathway
prior to the formation of porphyrinogens, ALA and PBG
will accumulate in the body tissues and fluids.
 These compounds can impair the function of abdominal
nerve and central nervous system, resulting in abdominal
pain and neuropsychiatric symptoms.
 This toxic effect of ALA and PBG may be due to
inhibition of ATPase in nervous tissue by ALA. ALA
may be taken up by brain which causes conduction
paralysis.
 On the other hand, enzyme blocks later in the pathway,
result in the accumulation of the porphyrinogens which
on exposure to light auto-oxidized to corresponding
porphyrin derivatives, and causes photosensitivity to
visible light and skin injuries.
 The injuries occur in the exposed areas where sunlight
activates porphyrins, which reacts with molecular
oxygen to form oxygen radicals in the skin.
 These oxygen radicals injure lysosomes and other
organelles. Damaged lysosomes release their degradative
enzymes, causing skin damage.
Figure 14.4: Biochemical basis of the major signs and
symptoms of porphyrias.
 In congenital erythropoietic porphyria the urine

of these patients is red because of excretion of large

amounts of uroporphyrinogen I and their teeth

exhibit a strong red fluorescence under ultraviolet

light because of the deposition of porphyrins.

 Furthermore, their skin is usually very sensitive to

light because of photosensitive porphyrins.

Figure 14.5: Fluorescent teeth of a patient with
porphyria cutanea tarda.
Figure 14.6: Skin lesion in a patient with porphyria
cutanea tarda.
Treatment of Porphyria
• Present treatment of porphyria is basically symptomatic.
 The symptoms of most porphyrias are now readily
controlled with dietary changes or administration of
heme or heme derivatives.
 Avoid drugs that cause production of cytochrome P450.
 Patients exhibiting photosensitivity may benefit from
administration of β-carotene.
 β-carotene decreases the production of free radicals and
helps to diminish photosensitivity.
 Use of sunscreens that filter out visible light can also be
helpful to decrease the photosensitivity.
 Ingestion of large amounts of carbohydrate and
administration of hematin to repress synthesis of ALA
synthase.
 ALA synthase diminish production of harmful heme
precursors .
Laboratory Test for the Detection of Porphyrins
and their Precursors
 Spectrophotometry is used to detect porphyrins and their
precursors.
 Coproporphyrins and uroporphyrins are excreted in urine
increased amounts in the porphyrias.
 When present in urine or feces, they can be separated by
extraction with appropriate solvents then identified and
quantified using spectrophotometric methods.
 After approximately 120 days, the heme group of
hemoglobin released from dying erythrocytes in the
spleen is degraded to yield free Fe 2+ and ultimately
bilirubin.
 First hemoglobin is dissociated into heme and globin.
 Globin is degraded to its constituent amino acids, which
are reused.
Thank you