CHACTERISTICS &

BIOSYNTHESIS OF
HEMOGLOBIN
Prepared by: EMNIEL BALBUENA BMLS 3-A
KATHLEEN BRAZAS BMLS 3-B
Turgeon 5th & 6th Edition
 Topic Coverage:
Characteristics & Biosynthesis of Hemoglobin

Genetic Inheritance of Hemoglobin

Chemical Composition and Configuration of Hemoglobin
The Role of 2,3 - Diphosphoglycerate
Oxygen Dissociation and Alterations
Carbon Dioxide Transport
Biosynthesis of Hemoglobin
Formation From Porphyrin
The Role of Iron in Hemoglobin Synthesis
Hepcidin
Globin Structure and Synthesis
Alpha Globin Locus
Beta Globin Locus
Felix Seyler
In 1862, he identifided the respiratory protein
hemoglobin and discovered the characteristic color
spectrum of hemoglobin .

Dissociation curve
a graph that represents the relationship between the
amount of oxygenbound to hemoglobin and the partial
pressure in oxygen in blood.
Genetic Composition and
Configuration of Hemoglobin
Normal adult hemoglobin A is inherited in simple
mendelian fashion
Phenotype A/A

approximately 350 variant types

Most defects in hemoglobin are related to either amino

acid substitutions or diminished production of one of the
polypeptide chains.
Disorders referred to as hemoglobinopathies
represent disorders related to defective
hemoglobin molecules.
Chemical Composition and
Configuration of Hemoglobin
Normal adult hemoglobin (hemoglobin A)
consists of four heme groups and four
polypeptide chains with a total of 574 amino
acids.

The polypeptide chains are organized into

two alpha chains and two beta chains
Structure of the hemoglobin
Chemical Composition and
Configuration of Hemoglobin
Normal adult hemoglobin has 141 amino
acids in each of the alpha chains and 146
amino acids in each of the beta chains.

The four hemes and four polypeptide chains are

assembled in
a very specific spatial configuration
The heme portion of the hemoglobin
molecule consistsconsists of 1 iron (Fe2+) Each of the four chains in the molecule coils
atom and 4 pyrrole rings
into eight helices, forming an egg-shaped
molecule with a central cavity
complete hemoglobin molecule
consists of 4 heme molecules
Chemical Composition and
Configuration of Hemoglobin
Metabolic processes within the erythrocyte ensure a suitable
intracellular environment for hemoglobin that protects it from
chemical changes that might result in the loss of its native
structure or denaturation

If hemoglobin is denatured, it loses its ability to

carry oxygen.
The role of 2,3
Diphospoglycerate

The major function of the hemoglobin

molecule is the transport of oxygen to
the tissues.
The manner in which 2,3-DPG binding to
reduced hemoglobin (deoxyhemoglobin)
affects oxygen affinity is complex.

, 2,3- DPG combines with the beta chains of

deoxyhemoglobin and diminishes the molecule’s affinity for
oxygen
The role of 2,3
Diphospoglycerate
When the individual heme groups
unload oxygen in the tissue, the beta
chains are pulled apart.

These activities result in a progressively lower

affi nity of the molecule for oxygen.

With oxygen uptake in the lungs, the salt bonds are

sequentially broken; the beta chains are pulled together.
The role of 2,3
Diphospoglycerate

In cases of tissue hypoxia, oxygen moves from

hemoglobin into the tissues, and the amount of
deoxyhemoglobin in the erythrocytes increases.

If hypoxia persists, depletion of free 2,3-DPG

leads to increased production of more 2,3-DPG
and a persistently lowered affnity of the
hemoglobin molecule for oxygen.
Oxygen Dissociation and
Alterations

The structure of the hemoglobin

molecule makes it capable of
considerable molecular changes as it
loads and unloads oxygen
Oxygen Dissociation

Figure 5.15
graphically describe the relationship between oxygen
content (percentage of saturation) and partial pressure In humans, the P50 value is 26.52 mm Hg for whole
of oxygen (PO2 ).
blood under accepted standard conditions of pH
7.4 and temperature of 37.5°C.
P50 value is defi ned as the partial pressure of oxygen required to
produce half saturation of hemoglobin, when the
deoxyhemoglobin (reduced hemoglobin) concentration equals the
oxyhemoglobin (oxygenated hemoglobin) concentration at a
constant pH and temperature
Oxygen Dissociation

Figure 15

The oxygen dissociation curve of normal adult blood. The oxygen tension at 50% oxygen
saturation (P50) is approximately 27 Torr. As the curve shifts to the right‚ the oxygen affi
nity of the hemoglobin decreases‚ and more oxygen is released at a given oxygen tension.
With a shift to the left‚ the opposite effects are observed. A decrease in pH or an increase in
temperature decreases the affinity of hemoglobin for oxygen.
Oxygen Dissociation

Oxygenation of one site on a

hemoglobin molecule enhances affinity
for oxygen at a different but chemically
identical site.

The first oxygen molecule binds to an alpha

chain, causing a change in the three- and the alignment of the chains to each
dimensional structure of that chain. other rapidly changes

The addition of a second oxygen to the other The 2,3-DPG is expelled from the molecule,
alpha chain produces a change in the resulting in increased oxygen affinity, and
molecular structure oxygen is added to the remaining beta chain
Oxygen Dissociation and
Alterations
Alteration
Oxygen dissociation as represented
Fetal hemoglobin (hemoglobin F) by the sigmoid curve can be shifted
has an increased affinity for to the right (decreased oxygen
oxygen affinity) by a decrease in pH (Bohr
effect)
Oxygen Dissociation and Alterations

Alterations

Oxygen dissociation as represented

by the sigmoid curve can be shifted
to the right (decreased oxygen
affinity) by a decrease in pH (Bohr
effect)
Oxyhemoglobin is a stronger
acid than deoxyhemoglobin.

Because deoxygenated hemoglobin is more

alkaline than is oxygenated hemoglobin, and an
alkaline pH stimulates glycolysis, 2,3-DPG
production is thereby increased

in turn, decreases molecular affinity for oxygen.

Carbon Dioxide Transport

The transport function of hemoglobin also

includes support for carbon dioxide transport
from the tissues to the lungs.

A portion of this iron is retained within the cell

for use or for storage in ferritin
Carbon Dioxide Transport
which accounts for approximately three fourths of the activity for removing carbon
dioxide, carbon dioxide diffuses into the erythrocytes, is catalyzed by the enzyme
carbonic anhydrase, and is transformed into carbonic acid.

The hydrogen ion of carbonic acid is accepted by the alkaline

deoxyhemoglobin, and the bicarbonate ion diffuses back into the
plasma.
Carbon Dioxide Transport
Chloride Shift or “Hamburger effect”
-Free bicarbonate diffuses out of
erythrocytes into the plasma in In the pulmonary
exchange for plasma chloride (Cl- ) that
capillaries, bicarbonate is
diffuses into the cell.
converted back into
carbon dioxide
and water and eliminated
Bicarbonate is carried back to
through respiration.
the lungs by the plasma.
Carbon Dioxide Transport
Approximately one fourth of the total This stabilizes the deoxy
carbon dioxide exchanged by formand decreases oxygen affi
erythrocytes in respiration is by a nity. Approximately 5% of
carbon dioxide is carried in
direct transport mechanism
solution in plasma to the lungs.
deoxyhemoglobin directly binds
with carbon dioxide.
This carbon dioxide reacts with uncharged
amino groups of the four globin chains to
form negatively charged carbamino
hemoglobin
Biosynthesis of Hemoglobin
Hemoglobin is synthesized during
most of the erythrocytic maturation
process.
65% of cytoplasmic hemoglobin is
synthesized before the nucleus is
extruded
35% is synthesized in the early reticulocyte.
Biosynthesis of Hemoglobin
Several types of hemoglobin can be found during
normal human growth and development:
A, A2, F and embryonic forms.

A - 2 alpha, 2 beta
A2 - 2 alpha, 2 delta
F - 2 alpha, 2 gamma

Normal adult hemoglobin A - predominant type of

hemoglobin. Genotype: A/A.
Biosynthesis of Hemoglobin

Genetic defects related either to amino acid

substitutions or to diminished production of one of
the polypeptide chains result in abnormal
hemoglobin molecules. These conditions are
referred to as hemoglobinopathies. One of the best-
known hemoglobinopathies is sickle cell anemia.

Hemoglobin S - normal adult hemoglobin A variant

Biosynthesis of Hemoglobin
Normal concentration of hemoglobin differs in children,
adult females, and male adults.

Various factors have an effect on the concentration of

hemoglobin. These factors are:
age
gender
pregnancy
altitude
smoking
associated disease
altered hemoglobin derivatives
(carboxyhemoglobin, methemoglobin,
sulfhemoglobin)
Biosynthesis of Hemoglobin
Formation of Heme
from Porphyrin
Heme synthesis occurs in
most body cells except for
mature erythrocytes.
The red
bone marrow and the liver are the most predominant
heme
(porphyrin) producers

Heme produced in the erythroid

precursors is chemically identical to that
in the cytochromes and myoglobin.
Heme Biosynthetic Pathway

1. Succinyl-coenzyme A (CoA) condenses with glycine. An unstable intermediate, adipic acid is

formed from this condensation and is readily decarboxylated to delta-aminolevulinic acid (ALA).
This initial condensation reaction occurs in the mitochondria and requires vitamin B6 . The most
important limiting step in this reaction is the rate of conversion to delta-ALA, which is catalyzed
by the enzyme ALA synthetase.
2. Following the formation of delta-ALA in the mitochondria, the synthesis reaction continues in
the cytoplasm.
3. Two molecules of ALA condense to form the monopyrrole porphobilinogen (PBG). This
reaction is catalyzed by the enzyme ALA dehydrate.
4. Four molecules of PBG condense into a cyclic tetrapyrrole to form uroporphyrinogen I or III.
The type III isomer is converted, by way of coproporphyrinogen III and protoporphyrinogen, to
protoporphyrin.
5. The final steps, carried out in the mitochondria, involve the formation of protoporphyrin and
the incorporation of iron to form heme. Four of the six ordinate positions of ferrous (Fe 2+ ) iron
are chelated to protoporphyrin by the enzyme heme synthetase ferrochelatase. This step
completes the formation of heme, a colored compound consisting of four pyrrole rings
connected by methene bridges into a larger tetrapyrrole structure.
Formation of Heme
from Porphyrin
The synthesis of heme is a complex process that
involves multiple enzymatic steps

STEPS:

The process begins in the mitochondrion with the

condensation of succinyl-CoA and glycine to form
5-aminolevulinic acid

In the final enzymatic steps, iron is inserted into the ring

structure of protoporphyrin IX to produce heme.
Formation of Heme
from Porphyrin
STEPS:

Four molecules of PBG condense into a cyclic

tetrapyrrole to form uroporphyrinogen I or III. The type
III isomer is converted, by way of coproporphyrinogen
III and protoporphyrinogen, to protoporphyrin.

The final steps, carried out in the mitochondria, involve the

formation of protoporphyrin and the incorporation of iron
to form heme.

Four of the six ordinate positions of ferrous (Fe2+) iron

are chelated to protoporphyrin by the enzyme heme
synthetase ferrochelatase
The Role of Iron in
Hemoglobin Synthesis
IRON: most abundant transition metal in the body.

The daily loss of iron, 1 to 2 mg in adults, represents less than 0.1% of the 3
to 4 g of total iron in the human body and must be replaced from dietary
sources to maintain iron balance.

Most of the iron of the body is in the form of hemoglobin in red cells -
contain about 1 mg of iron per mL of RBCs or about 2 to 3 g of iron total.

Blood plasma contains only 2 to 3 mg of iron.

Transferrin - the plasma iron carrier that is the exclusive source of iron in
erythropoiesis.
The Role of Iron in
Hemoglobin Synthesis
In the duodenum, dietary free iron is reduced to ferrous (+2) iron and taken up
from the intestinal lumen into the enterocytes by the iron transport protein
divalent metal transporter 1 (DMT1).

Iron may be stored as ferritin in the enterocytes or exported into the

circulation by another iron transport protein, ferroportin 1 (fpn1).

Ferroportin exports iron into plasma from absorptive enterocytes, from

macrophages that recycle the iron of senescent erythrocytes, and from
hepatocytes that store iron.
Biopsy interpretation
of the liver
hemosiderin on H&E
stain.

Iron deposits are seen

as brown granules in
the hepatocyte
cytoplasm.
Beta Globin Locus
The genes are arranged sequentially.
The sequence of the genes is epsilon, gamma, delta, and beta.
There are two copies of the gamma gene on each chromosome 11.
The others are present in single copies.
Each cell has two beta globin genes, one on each of the two chromosomes 11
in the cell.
The polypeptide chains of globin are produced, as are other body proteins,
on the ribosomes. The alpha polypeptide chain unites with one of three other
chains to form a dimer and ultimately a tetramer.
Alpha Globin Locus
Each chromosome 16 has two alpha globin genes that are
aligned one after the other on the chromosome.
The two alpha globin genes are identical.
Each cell has a total of four alpha globin genes.
Each of the four genes produces about one quarter of the
alpha globin chains needed for hemoglobin synthesis. The
mechanism of this coordination is unknown.
The transiently expressed embryonic genes that substitute
for alpha very early in development, designated zeta, are also
in the alpha globin locus.
Summary of the Assembly of a
Hemoglobin Molecule

A hemoglobin molecule is assembled in sequential steps. These steps are as

follows:
1. The α- and β-globin polypeptides are translated from appropriate mRNAs.
2. Once heme containing iron binds to each of the four polypeptide chains,
the protein folds into a native 3-D structure.
3. The α and β units bind to each other by electrostatic bonding.
4. An unstable intermediate encounter complex can rearrange the units to
form a stable αβ dimer.
5. Two dimers combine to form the functional hemoglobin molecule, an α2 β2
tetramer.
Thank you!