Review on hemoglobin.

• Hemoglobin (Hb) is one of the most studied proteins in the body due to the
ability to easily isolate it from red blood cells (RBCs).
• Free (non-RBC) hemoglobin, generated from RBCs through hemolysis, has
a short half-life outside of the RBCs. When released into the plasma, it is
rapidly salvaged to preserve its iron and amino acid components; when
salvage capacity is exceeded, it is excreted by the kidneys.
• Hemoglobin’s main function is to transport oxygen from the lungs to tissues
and transport carbon dioxide from the tissues to the lungs for exhalation.
• Hemoglobin also contributes to acid-base balance by binding and releasing
hydrogen ions and transports nitric oxide (NO), a regulator of vascular tone.
Hemoglobin
structure
Hemoglobin is the first
protein whose structure
was described using x-ray
crystallography. The
hemoglobin molecule is a
globular protein consisting
of two different pairs of
polypeptide chains and four
heme groups, with one
heme group imbedded in
each of the four
polypeptide chains https://images.app.goo.gl/yiqpJ9PMNDexmUk
77
Heme Structure
Heme consists of a ring of carbon,
hydrogen, and nitrogen atoms called
protoporphyrin IX, with a central atom
of divalent ferrous iron (Fe2+). Each of
the four heme groups is positioned in a
pocket of the polypeptide chain near the
surface of the hemoglobin molecule.
The ferrous iron in each heme molecule
reversibly combines with one oxygen
molecule. When the ferrous irons are
oxidized to the ferric state (Fe3+),
they no longer can bind oxygen.
Oxidized hemoglobin is also called
methemoglobin. https://images.app.goo.gl/yUZUXR5k63vymM
wJ8
Globin structure

• The four globin chains comprising each hemoglobin

molecule consist of two identical pairs of unlike
polypeptide chains, 141 to 146 amino acids each.
Variations in amino acid sequences give rise to different
types of polypeptide chains. Each chain is designated by
a Greek letter.
Each globin chain is
divided into eight helices
separated by seven
nonhelical segments. The
helices, designated A to H,
contain subgroup
numberings for the
sequence of the amino
acids in each helix and are
relatively rigid and linear.
Complete hemoglobin
molecule
The hemoglobin
molecule can be
described by its
primary, secondary,
tertiary, and
quaternary protein
structures.
https://images.app.goo.gl/T2TcWmgoAn6RnhWC
A
• The primary structure refers to the amino acid sequence
of the polypeptide chains.
• The secondary structure refers to chain arrangements in
helices and nonhelices.
• The tertiary structure refers to the arrangement of the
helices into a pretzel-like configuration.
The quaternary structure of
hemoglobin, also called a
tetramer, describes the
complete hemoglobin
molecule.
The complete hemoglobin
molecule is spherical, has four
heme groups attached to four
polypeptide chains, and may
carry up to four molecules of
oxygen. The predominant adult
haemoglobin, HbA.
https://images.app.goo.gl/dP5xBu99VhXfRKAT6
• Hb A, is composed of two α-globin chains and two β-globin
chains. Strong α1–β1 and α2–β2 bonds hold the dimers
in a stable form.
– The α1–β2 and α2–β1 bonds are important for the stability of
the quaternary structure in the oxygenated and
deoxygenated forms.
HbA1c
• A small percentage of Hb A is glycated. Glycation is a posttranslational
modification formed by the nonenzymatic binding of various sugars to
globin chain amino groups over the life span of the RBC.
• The most characterized of the glycated hemoglobins is Hb A1c, in which
glucose attaches to the N-terminal valine of the β chain. Normally, about
4% to 6% of Hb A circulates in the A1c form.
• In uncontrolled diabetes mellitus, the amount of A1c is increased
proportionally to the mean blood glucose level over the preceding 2 to 3
months.

https://images.app.goo.gl/44eD2u6suoJrW3KG9
Hemoglobin biosynthesis
Heme biosynthesis

• Heme biosynthesis occurs in the mitochondria and

cytoplasm of bone marrow erythrocyte precursors,
beginning with the pronormoblast through the
circulating polychromatic (also known as
polychromatophilic) erythrocyte. As they lose their
ribosomes and mitochondria (location of the
citric/tricarboxylic acid cycle), mature erythrocytes can
no longer make hemoglobin.
https://images.app.goo.gl/Rhyxj3wtyYbZqJ9z5
• Heme biosynthesis begins in the mitochondria with the condensation of
glycine and succinyl coenzyme A (CoA) catalyzed by aminolevulinate
synthase to form aminolevulinic acid (ALA).
• In the cytoplasm, aminolevulinic acid dehydratase (also known as
porphobilinogen synthase) converts ALA to porphobilinogen (PBG). PBG
undergoes several transformations in the cytoplasm from
hydroxylmethylbilane to coproporphyrinogen III. This pathway then
continues in the mitochondria until, in the final step of production of
heme, Fe2+ combines with protoporphyrin IX in the presence of
ferrochelatase (heme synthase) to make heme.
https://images.app.goo.gl/aMGV6perWehvSCxMA
• Transferrin, a plasma protein, carries iron in the ferric (Fe3+)
form to developing erythroid cells.
– Apotransferrin- unbound to iron
• Transferrin binds to transferrin receptors on erythroid precursor
cell membranes and the receptors and transferrin (with bound
iron) are brought into the cell in an endosome.
• Acidification of the endosome releases the iron from
transferrin.
• Iron is transported out of the endosome and into the
mitochondria where it is reduced to the ferrous state, and is
united with protoporphyrin IX to make heme.
• Heme leaves the mitochondria and is joined to the globin
chains in the cytoplasm.
https://images.app.goo.gl/WZFqZYwzDChmfEgLA
Globin biosynthesis
• Six structural genes code for six globin chains. The α- and ζ-globin
genes are on the short arm of chromosome 16; the ε-, γ-, δ-, and β-
globin gene cluster is on the short arm of chromosome 11.
• In the human genome, there is one copy of each globin gene per
chromatid, for a total of two genes per diploid cell, with the exception of α
and γ. There are two copies of the α- and γ-globin genes per chromatid,
for a total of four genes per diploid cell.
https://images.app.goo.gl/BAsu2LTHoqgWpN3p6
• The production of globin chains takes place in erythroid
precursors from the pronormoblast through the circulating
polychromatic erythrocyte, but not in the mature erythrocyte.
• Transcription of the globin genes to messenger ribonucleic
acid (mRNA) occurs in the nucleus, and translation of mRNA
to the globin polypeptide chain occurs on ribosomes in the
cytoplasm.
• Although transcription of the α-globin genes produces more
mRNA than the β-globin gene, there is less efficient translation of
the α-globin mRNA. Therefore, the α and β chains are
produced in approximately equal amounts. After translation is
complete, the chains are released from the ribosomes in the
cytoplasm.
Hemoglobin assembly
After their release from ribosomes, each globin chain binds to a heme molecule,
then forms a heterodimer. The non-α chains have a charge difference that
determines their affinity to bind to the α chains. The α chain has a positive
charge
and has the highest affinity for a β chain due to its negative charge. The γ-globin
chain has the next highest affinity, followed by the δ-globin chain. Two
heterodimers
then combine to form a tetramer. This completes the hemoglobin molecule.
Two α and two β chains form Hb A, the major hemoglobin
present from 6 months of age through adulthood.
Hb A2 contains two α and two δ chains. Consequently, Hb
A2 comprises less than 3.5% of total hemoglobin in adults.
Hb F contains two α and two γ chains. In healthy adults, Hb
F comprises 1% to 2% of total hemoglobin, and it is present
only in a small proportion of the RBCs (uneven distribution).
These RBCs with Hb F are called F or A/F cells.
https://images.app.goo.gl/WeHhnP1GrrM6rdES8