STRUCTURE OF MYOGLOBIN AND HEMOGLOBIN

FLORENCE RAJARETNAM M.Sc., M.Phil

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Myoglobin and Hemoglobin:
 Myoglobin and hemoglobin carry out their biol
ogical functions by selectively and reversibly
binding molecules such as molecular oxygen o
r carbon dioxide.

 They belong to a group of proteins called hem

eproteins

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Structure and function of Myoglobin (Mb).

• Myoglobin hemeprotein
• location primarily in heart and cytosol of
skeletal muscle
• functions as a reservoir for oxygen and
as an oxygen carrier that increases the rate of tr
ansport of oxygen within the muscle cell.

 The stored oxygen is readily available to orga

nelles particularly in the mitochondrion where o
xidative metabolism is carried out.

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• Myoglobin comprises a single polypeptide chain
of 153 amino acids with a compact spherical stru
cture.

• The interior portion of the myoglobin molecule i

s composed almost entirely of nonpolar amino aci
ds forming a structure stabilized by hydrophobic
interactions.

• charged amino acids are located almost exclus

ively on the surface of the myoglobin, where they
can form hydrogen bonds with water.

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•The heme group of myoglobin sits in a crevice in
the protein.

• This cavity is lined with nonpolar amino acids e

xcept for two histidine residues.

• One of them, termed the proximal histidine, bind

s directly to the iron of heme.

• The second or distal histidine does not directly

interact with the heme groups.

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The oxygen directly binds with the iron and forms
a hydrogen bond with the distal histidine.

STRUCTURE OF THE HEME GROUP

• Heme is a complex of protoporphyrin IX and ferr
ous iron (Fe+2).

•The iron is held in the center of the heme molecu

le by bonds to the four nitrogen of the porphyrin ri
ng.

•The (Fe+2) of heme can form two additional bond

s, one on each side of the planar porphyrin ring.

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• In myoglobin and hemoglobin, one position is co
ordinated to a histidine ( proximal) of the protein,
whereas the other position is available to bind ox
ygen.
• In deoxyhemoglobin, the sixth coordination site
remains unoccupied.

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STRUCTURE AND FUNCTION OF HEMOGLOB
IN
• Hemoglobin is found exclusively in the red blood c
ells, where its main function is to transport oxygen f
rom the lungs to the capillaries of the tissues.

• Hemoglobin A, the major hemoglobin in adults, co

mprises four polypeptide chains (two α-chains and t
wo β-chains, α2β2) held together by noncovalent inte
ractions.

• Each subunit has a helical structure and heme-bin

ding pocket similar to that described for myoglobin.

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• In foetus, the Hb is specifically called HbF
( foetal haemoglobin)- consisting of 2 alpha and 2
gamma chains and is different from adults.
• In adults , there are 3 different type of Hb ,
1) H b A ( 97 %)- major.
2) Hb A 2( 2%)- consisting of 2 alpha and 2 delta c
hains and
3) Hb F( 1 %)- consisting of 2 alpha and 2 gamma
chains.
• Alpha chain has 141 amino acids and beta and d
elta chains have 146 amino acids each.

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• The hemoglobin tetramer comprises of two identi
cal dimers, α1β1 and α2β2 (where the numbers refer
to dimer 1 and dimer 2).

• The two polypeptide

chains within each dimer
are held tightly together.

•The two dimers are able

to move with respect to
each other.

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• The two dimers occupy different positions in de
oxyhemoglobin and oxyhemoglobin.

•The deoxy-form of hemoglobin is called the T- or

taut form.

•The α1β1 and α2β2 dimers interact through a ne

twork of ionic bonds and hydrogen bonds that limi
t the movement of the polypeptide chains.

•The T-form is the low oxygen affinity form of he

moglobin.

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Oxygen binding to the Fe of heme

•The proximal histidine binds to the Fe on one side

of the porphyrin ring.

• O2 binds to Fe on the other side.

•On oxygenation, the iron atom which was slightly a

bove the plane of the heme, moves into the plane, p
ulling the proximal histidine with it.

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•This movement cleaves some of the salt links, an
d the equilibrium is shifted from T to R.

•This leads to a structure called the R or relaxed f

orm in which the dimers have more freedom of mo
vement.

•The R-form is the high oxygen affinity form of he

moglobin. 13
BINDING OF OXYGEN TO HEMOGLOBIN

• Hemoglobin can bind one oxygen molecule (O

2) to each of its four-heme-containing subunit
s.

• The saturation of these oxygen-binding sites (

Y) is defined as the fractional occupancy of t
he total binding sites.

• The value of Y can vary between zero (all site

s empty) and 100% (all four sites full)

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A plot of Y measured at different partial pressures
of oxygen is called the oxygen saturation curve or
oxygen-hemoglobin dissociation curve.

The curves for myoglobin and hemoglobin show i

mportant differences, which can be clearly observ
ed in Figure 2.

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Figure 2
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 Myoglobin has a higher oxygen affinity than h
emoglobin.

The oxygen dissociation curve for myoglobin h

as a hyperbolic shape.

The oxygen dissociation curve for hemoglobin

is sigmoidal in shape indicating that the subunit
s cooperate in binding oxygen.

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Cooperative binding of oxygen to H
emoglobin
• binding of an oxygen molecule at one heme in
creases the oxygen affinity of the remaining he
me groups in the same hemoglobin molecule (t
his effect is known as heme-heme interaction).

 Although it is difficult for the binding of the fi

rst oxygen molecule, subsequent binding of oxy
gen occurs with a high affinity .

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The cooperative binding of oxygen allows hemogl
obin to deliver more oxygen to the tissues in resp
onse to relatively small changes in the partial pre
ssure of oxygen.

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• In the lung, the concentration of oxygen is
high, and hemoglobin becomes virtually sa
turated (or "loaded") with oxygen (Y = 97%)
.

• In peripheral tissues oxyhemoglobin releas

es (or "unloads") much of its oxygen (Y = 5
0%) for use in the oxidative metabolism of
the tissues

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 Allosteric Effect of Hemoglobin molecul
e
• Allosteric effectors are small molecules that
bind to proteins
– at sites that are spatially different from the
ligand-binding sites.

• An allosteric effector may exert either:

– a positive influence on ligand interaction (i
ncreased affinity) or
– a negative influence (decreased affinity) .

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The ability of hemoglobin to bind oxygen rev
ersibly is affected by the following allosteric
effectors
1-Oxygen molecule:
 the binding of an oxygen molecule at one he
me group increases the oxygen affinity of the
remaining heme groups leading to the formati
on of the R-form with high oxygen affinity

 2-Carbon Dioxide: Some of the carbon dioxide

produced during metabolism is carried bound
to hemoglobin.

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• Approximately 80% of the CO2 produced in metabo
lizing cells is transported to the lungs in the form of
bicarbonate.
This transport process is referred to as
isohydric transport.

• Metabolizing cells produce CO2 which diffuses into

the blood and enters the circulating red blood cells (
RBCs).

•Within RBCs the CO2 is rapidly converted to carbon

ic acid through the action of
carbonic anhydrase

CO2 + H2O ——> H2CO3 ——> H+ + HCO3–

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 The bicarbonate ion produced in this dissociation
reaction diffuses out of the RBC and is carried in
the blood to the lungs.
.

A.In the tissues, CO2 is re

leased.
• In the RBC, this CO2 for
ms carbonic acid, w/c re
leases protons.
• The protons bind to oxy
genated Hb, causing it t
o release O2 to the tiss
ues

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B. In the lungs, the rxn’s are reversed.
• O2 binds to protonated Hb, causing the relea
se of protons.
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3-Hydrogen / proton(H+ )
• The hydrogen ions
produced in peripheral
tissues during metabolism
bind to hemoglobin.
• The binding of hydrogen
ion to the hemoglobin
molecule decreases
its affinity for oxygen.

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4 – 2,3-Bisphosphoglycerate (2,3BPG):
• 2,3BPG is synthesized from 1,3 BPG by the ac
tion of the enzyme Biphospho Glyceromutase.

• A molecule of 2,3BPG binds to a pocket in the

center of the deoxyhemoglobin tetramer.

• This preferential binding stabilizes the T- conf

ormation of Hemoglobin.

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• Thus the binding of a molecule of oxygen increa
ses the affinity of Hemoglobin by other oxygen
molecules,

• while the carbon dioxide, Hydrogen ion and 2,3

BPG, once bound to the hemoglobin molecule le
ad to a decrease in the affinity of Hemoglobin to
Oxygen.

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 BOHR EFFECT

• The H+ and C02 promote the release of 02 from

hemoglobin, an effect that is physiologically im
portant in enhancing the release of 02 in metab
olically active tissues such as muscle.

• Conversely, 02 promote the release of H+ and C0

2 in the alveolar capillaries of the lungs .
These allosteric linkages between the binding of
H+, C02 and 02 are known as the Bohr Effect

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 CARBON MONOXIDE AS A POISON

Carbon monoxide can bind the hemoglobin

molecule with more affinity than oxygen and i
nterferes in the oxygen transport to the tissue
s.

 The affinity of the isolated heme group is 2

5000 greater for CO than Oxygen, but this affi
nity is considerably reduced when the heme g
roup is incorporated into Myoglobin or Hemogl
obin.

 In a normal person, only 1% of the oxygen b

inding sites of Hemoglobin are occupied by C
O, Which is easily tolerated by the organism.

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