HEMOGLOBIN

METABOLISM
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• Hemoglobin is a conjugated
globular protein.
• Concentration in each RBC=34 g/dL.
• About 65% of Hgb synthesis occurs
during nucleated stages of RBC
maturation, and 35% occurs during
the reticulocyte stage.
• Vehicle for transport of O2

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HEMOGLOBIN
STRUCTURE

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• Normal hemoglobin consists
of globin (a tetramer of two
parts of unlike globin
polypeptide chains)
• four heme groups, each of
which contains a
protoporphyrin ring plus
iron. 
• one molecule of 2,3-DPG
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• Heme: (red)
– The iron-containing protoporphyrin
IX(carbon, hydrogen and nitrogen)
– iron is in the ferrous (Fe2+) state
• Globin:
– 2 pairs of polypeptide chains (141-146
AA)
– There are 4 globinheyzgran
chains in the 5
 Hemoglobin Synthesis
• Depends on three processes: 
1. adequate supply and
delivery of iron;
2. adequate synthesis of
protoporphorins (heme
precursor);
3. adequate globin synthesis
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heyzgran 7
 Hemoglobin Synthesis
• Occurs in the BM
• Mitochondria and cytoplasm from pronormoblast
to reticulocyte
1. Condensation of glycine and succinyl coenzyme
A (CoA) catalyzed by aminolevulinate synthase
(ALAS) to form aminolevulinic acid (ALA).
2. ALA dehydratase with ALA catalyzes the
formation of PORPHOBILINOGEN
3. Porphobilinogen deaminase hydroxyl
methylbilane
4. FINAL STEP= PRODUCTION OF HEME(Fe2+ +
ProtoporphyrinIX)
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 Iron Delivery and Supply 1
of 3
• Iron delivered to
membrane of RBC
precursor by protein
carrier transferrin.
• Most of the iron that
crosses membrane and
enters cytoplasm of cell
is committed to
hemoglobin synthesis. 
Proceeds to
mitochondria for heyzgran 9
 Iron Delivery and Supply 2
of 3

• Excess iron in cytoplasm

aggregates as
FERRITIN. 
• Amount of ferritin stored
depends on ratio
between level of plasma
iron and amount of iron
required by erythrocyte
for hemoglobin synthesis.
• Two-thirds of total iron
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 Iron Delivery and Supply
3 of 3

• Sideroblast:
– A ferritin-containing normoblast
(nucleated RBC) in the bone marrow.

– It makes up from 20% to 90% of

normoblasts in the marrow.
• Siderocyte:
– A nonnucleated red blood cell
containing iron in a form other than
hematin.
– Confirmed by a specific iron stain
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 Synthesis of
Protoporphyrins 3 of 3

• At end of heme synthesis,

small amount of excess
porphyrin in mitochondria
is complexed to zinc. 
Excess is called free
erythrocyte
protoporphyrin (FEP). 
FEP is elevated when iron
supply depleted.
• HEME leaves the
mitochondria and is joined
in the globin chains in the
cyplasm heyzgran 12
Globin Synthesis
• Occurs on RBC-specific ribosomes
which are derived from inheritance
of various structural genes. 
• Each RBC contains four alpha, two
zeta, two beta, two delta, two
epsilon, and four gamma genes. 
• Resulting gene products are alpha,
zeta, beta, delta, epsilon, and
gamma globin chains.
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Hemoglobin Greek Symbol
Chain
Alpha α
Beta β
Delta δ
Epsilon ε
Gamma γ
Zeta ζ
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• Throughout embryonic and fetal
development, activation of globin genes
progresses from zeta to alpha, and from
epsilon to gamma to delta to beta.

zeta alpha

epsilon gamma delta beta

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 Hemoglobin Globin Chain
Types
Gower I 2 zeta and 2 epsilon
Gower II 2 alpha and 2 epsilon
Portland 2 zeta and 2 gamma
Hemoglobin F 2 alpha and 2 gamma
(fetal Hgb)
Hemoglobin A 2 alpha and 2 beta
(Adult Hgb)

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heyzgran 17
• Epsilon and zeta globins usually
appear only during embryonic
development.  Gower I Hemoglobin
is two zeta and two epsilon chains. 
Gower II Hemoglobin is composed
of two alpha and two epsilon chains. 
Hemoglobin Portland is composed
of two zeta and two gamma chains.

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• Beta chain production steadily
increases after birth until adult
percentages are reached between
three months and six months of
age.
• All normal adult hemoglobins
(Hemoglobin A) consists of two
alpha and two non-alpha globin
chains.
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• Hemoglobin A has two alpha and
two beta chains and comprises
95-97% of adult hemoglobin. 
Hemoglobin A2 has two alpha and
two delta chains and comprises 2-
3% of total adult hemoglobin.

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• Each globin chain links
with heme molecule
(protoporphyrin ring
with iron) to form
hemoglobin.
• Entire hemoglobin
molecule has two alpha
chains, two beta chains,
and four heme groups.
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• Rate of globin synthesis directly
related to rate of porphyrin
synthesis and vice versa. 
• Iron accumulates in RBC
cytoplasm as ferritin aggregates. 
Iron-laden RBC called sideroblast
and anucleated from called
siderocyte.  (Will see iron
clusters when cells stained with
Prussian Blue stain).
• Completed hemoglobin molecule
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 2,3-
Diphosphoglycerate
(2,3-DPG)
• Is an organic phosphate
responsible for hemoglobin's
affinity for oxygen.
• Is a product derived from
Luebering-Rapoport shunt.
• Is located in the central cavity of
hemoglobin molecule.  Is bound to
beta chains. heyzgran 23
 Assembly of Hemoglobin
Molecule
1 of 2

• Formation of hemoglobin requires

iron, globin chains, protoporphyrin
IX, and 2,3-DPG.
• To assemble molecule, ferric iron
(Fe3+) must be obtained from
ferritin.  Iron is chemically reduced
(Fe2+), and then inserted as
ferrous iron into center of
protoporphyrin IX molecule.
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 Assembly of Hemoglobin
Molecule
2 of 2

• When globin chain completed on

ribosome, it is released to cytoplasm.
Individual alpha and beta chains
quickly and spontaneously form alpha-
beta dimers.  Two heme molecules
bind to each alpha-beta dimer.  Two
dimers quickly form a tetramer and
assume final three dimensional shape.
• Last step is insertion of 2,3-DPG
molecule into center cavity of
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heyzgran 26
Hemoglobin
Function

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• Primary function is delivery and
release of oxygen to tissues. Each of 4
heme iron atoms can bind 1 oxygen
molecule.
• 1.34 mL of Oxygen is bound/1 g of Hb
• One of most important controls of
hemoglobin affinity for oxygen is RBC
organic phosphate: 2,3-
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• Unloading of oxygen by
hemoglobin accompanied by
widening of space between
beta chains and binding of 2,3-
DPG to beta chains. 
• Resulting hemoglobin molecule
known as "tense form“, which
has a lower oxygen affinity. 
Also known asheyzgran 29
Tense Form

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• When hemoglobin binds
oxygen, 2,3-DPG and beta
chain bonds break.  Beta
chains close up and 2,3-DPG
expelled.  Is "relaxed form" of
hemoglobin.  Has a high
affinity for oxygen.  Is also
called oxyhemoglobin.
• Conversion between tense
form and relaxed form referred
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Relaxed Form

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heyzgran 33
• Relationship between
hemoglobin and oxygen has a
sigmoid curve shape - is called
the oxygen dissociation
curve. 
• Shape of curve means lots of
oxygen can be delivered to
tissues with small drop in
oxygen tension.
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Oxygen Dissociation Curve

% Sat

Tissue pO2
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Oxygen Dissociation Curve

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• Normally, PO2=27mmHg
results in 50% oxygen
saturation of Hb molecule.

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• In lungs, where pO2 (oxygen
tension) very high,
hemoglobin almost 100%
saturated with oxygen.  As
RBCs travel to tissues where
oxygen tension drops,
hemoglobin saturation drops
to about 75%, releasing
oxygen to tissues.
• Is normally occurring process.
heyzgran 38
• In hypoxia, a compensatory
"shift to right" of
hemoglobin dissociation
curve occurs to relieve tissue
oxygen deficit.  Right shift of
curve, mediated by increase
in 2,3-DPG, results in
decrease in hemoglobin's
affinity for oxygen and an
increase in oxygen delivery to
tissues.
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• In "shift to left“, 50% oxygen
saturation og Hb occurs at a PO2
less than 27mm Hg.
• In “shift of the curve to the right”,
50% oxygen saturation of Hb
occurs at PO2 higher than 27 mm
Hg.
• In "shift to left“, higher % oxygen
saturation and high oxygen
affinity.
• In “shift of the curve to the right”,
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• In "shift to left" of hemoglobin
dissociation curve, see an
increase in hemoglobin-oxygen
affinity and decrease in amount of
oxygen being delivered to tissues.
• Conditions causing a "shift to
left" include alkalosis, increase in
the amount of abnormal
hemoglobins (methemoglobin or
carboxyhemoglobin), increased
amount of Hemoglobin F, or
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• Hemoglobin-oxygen affinity
also expressed by P50 values. 
P50 is point at which
hemoglobin is 50% saturated
with oxygen.  Increase in P50
values represents  a decrease
in hemoglobin-oxygen affinity
(shift to right).  Decrease in
P50 values represents increase
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• CARBON DIOXIDE TRANSPORT
• Venous blood-CO2 diffuses
into RBC combines with H2O
to form carbonic acid
(H2CO3)- carbonic
anhydrase=release H+ and
bicarbonate (HCO3¯)
• CO2 expelled in the lungs.
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• NITRIC OXIDE TRANSPORT
• Nitric oxide-secreted by
vascular EC-causes relaxation
of the vascular wall smooth
muscle and vasodilation.
Some enters RBC.

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 DYSHEMOGLOBINS
• Dysfunctional Hbs-unable to
transport oxygen)
• Methemoglobin-reversible
oxidation of heme iron to
ferric state
• Sulfhemoglobin-irreversible
oxidation of Hb by drugs
• Carboxyhemoglobin-
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End…
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