Hemoglobin and

Insulin
Characterization
1. Hemoglobin
Overview of Hemoglobin

• Hemoglobin (Hb)= Globin + Heme

• Hemoglobin is a protein found exclusively in red blood

cells (RBCs), where its main function is to transport
oxygen (O2) from lungs to capillaries of tissues.
Structure of hemoglobin
• Hemoglobin A, the major hemoglobin in
adults, is composed of four polypeptide
chains: two α chains and two β
chains, held together by noncovalent
interactions.

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 Structure of
hemoglobin
• The alpha globin chain is
composed of 141 amino acids
& the beta globin chain is
composed of 146 amino acids

• Both alpha and beta proteins

share similar secondary and
tertiary structures
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Each hemoglobin molecule (globular protein) consist of 4 Heme and
4 polypeptide chains each one bind to a Heme.
 Heme
• Heme is a complex of protoporphyrin IX and
ferrous iron (Fe2+).

• Heme = protoporphyrin + Iron

• The iron is held in the center of heme molecule

by bonds with four nitrogen atoms of
protoporphyrin rings.
 Heme
• (Heme Fe2+) can form two additional bonds, one on each side of
the protoporphyrin ring:

• One is bonded to the side chain of a histidine residue within the

globin molecule

• The second is available to bind oxygen or CO2

• Hence, in hemoglobin, one of these positions is coordinated to

the side chain of a histidine residue of the globin molecule,
whereas the other position is available to bind oxygen.
 Quaternary structure of hemoglobin
• The hemoglobin tetramer is composed of two identical dimers, (αβ)1
and (αβ)2, in which the numbers refer to dimers one and two. Bonds
strongest Covalent :
Peptide Disulfide
Noncovalent : 2nd

The two polypeptide chains within each dimer are tightly held
strongest non-polar :
• hydrophobic
weakest: polar : ionic
h-bond

together, primarily by hydrophobic interactions

Quaternary structure of hemoglobin
• In contrast, the two dimers are able to move with respect to
each other, being held together primarily by polar bonds (H
or ionic bond).
• The weaker interactions between these mobile dimers result in
the two dimers occupying different relative positions in
deoxyhemoglobin as compared with oxyhemoglobin
Quaternary structure of hemoglobin

• T form: The deoxy form of hemoglobin is called the “T,” or taut (tense)
form. In the T form, the two αβ dimers interact through a network of ionic
bonds and hydrogen bonds that limit the movement of the polypeptide
chains. The T form is the low oxygen- affinity form of hemoglobin.

• R form: The binding of oxygen to hemoglobin causes the rupture of some

of the ionic bonds and hydrogen bonds between the αβ dimers. This leads
to a structure called the “R,” or relaxed form, in which the polypeptide
chains have more freedom of movement. The R form is the high oxygen-
affinity form of hemoglobin.
 Binding of oxygen to hemoglobin
• hemoglobin can bind to four oxygen molecules —one at each of its four heme groups.

• The degree of saturation (Y) of these oxygen-binding sites on all hemoglobin

molecules can vary between zero (all sites are empty) and 100% (all sites are full).

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Oxygen Cooperative Binding
• Cooperative binding of oxygen to the four
subunits of hemoglobin means that the
binding of an oxygen molecule at one heme
group increases the oxygen affinity of the
remaining heme groups in the same
hemoglobin molecule.
 Oxygen Dissociation Curve
• A plot of (Y) measured at different partial pressures of oxygen
(pO2) is called as the oxygen dissociation curve.

• Hemoglobin bind to O2 at high O2 tension (pressure) in the lung

and delivered it to the tissues at low O2 tension (pressure).

• –The oxygen binding is cooperative: high pressure more oxygen

• As each O2 binds to hemoglobin, the molecule undergoes a

conformational change increasing the O2 affinity for the
remaining subunits. This creates the sigmoidal oxygen
dissociation curve
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 2,3 - Bisphosphoglycerate
• Allosterically regulates Oxygen binding to hemoglobin, BPG is present in relatively high
concentrations in Erythrocytes.

• 2,3-Bisphosphoglycerate is known to greatly reduce the affinity of hemoglobin for oxygen—

there is an inverse relationship between the binding of O2 and the binding of BPG.

• Only one molecule of BPG is bound to each hemoglobin tetramer. BPG lowers hemoglobin’s
affinity for oxygen by stabilizing the T state. oxygen won't bind
For general
knowledge
Question

Relaxed Oxyhemoglobin taut deoxyhemoglobin

2-3 dpg is present
 Types of hemoglobin
• HBA: the major hemoglobin in humans

• HBF: normally synthesized only during fetal

development

• HBA2: first appears 12 weeks after birth- a

minor component of normal adult HB

• HBA1C : has glucose residues attached to β-

globin chains – increased amounts in
Diabetes Mellitus
Think…

• Why muscles appear as red-brown in

color?

• Why some mammals (such as

whales) are able to remain
submerged for long periods?
Myoglobin

• myoglobin, a protein found in the muscle or cardiac

cells of animals. It functions as an oxygen-storage unit,
providing oxygen to the working muscles as necessary.

• Functions both as a reservoir for oxygen, and as an

oxygen carrier that increases the rate of transport of
oxygen within the muscle cell.

• Myoglobin consists of a single polypeptide chain.

Related Disease
 Sickle Cell Anemia
(hemoglobin S disease)
• Sickle cell anemia is a genetic disorder of the blood caused by a
single alteration in the gene for β-globin.

• The lifetime of an erythrocyte in sickle cell anemia is less than 20

days, compared with 120 days for normal RBCs.

• Sickle cell anemia is characterized by lifelong pain, chronic

hemolytic anemia with associated hyperbilirubinemia, and
increased susceptibility to infections.

• An infant does not begin showing symptoms of the disease until

sufficient Hb-F has been replaced by Hb-S .
 Thalassemias
• The thalassemias are hereditary hemolytic diseases in which
an imbalance occurs in the synthesis of globin chains.

• Normally, synthesis of the α- and β-globin chains is

coordinated, so that each α-globin chain has a β-globin chain
partner. This leads to the formation of α2β2 (Hb A).

• In the thalassemias, the synthesis of either the α- or the β-

globin chain is defective.
2. Insulin
 Insulin:
• Insulin is a hormone central regulating
carbohydrate and fat metabolism in the
body.

• Insulin causes liver cells, muscle cells

and fat tissue to take up glucose from
the blood and store it as glycogen in the
liver and muscle. increase glycogenesis

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Production of insulin:
• The pancreas has two important functions :
1. Producing hormones – such as insulin and glucagon which
regulate blood sugar levels.
2. Producing pancreatic digestive enzymes.

• Insulin is released when any of the several stimuli are detected–

stimuli include ingested protein and glucose in the blood from
digested food.

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 Insulin structure
• Insulin is produced and stored in the body as a hexamer
(preprohormone), while the active form is the monomer.

• Active insulin is a peptide hormone composed of 51 amino

acids arranged in two chains (A and B) and has a molecular
weight of 5808 Da.

• Mature insulin consists of the 21 amino acids in A-chain, and

30 amino acids in B-chain linked together by disulfide bonds.
tertiary
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Amino acids Sequence of Insulin

Insulin is synthesized as a preprohormone 31

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 Actions of insulin on the human
metabolism

• Control of the cellural intake of certain substances.

• Regulation of DNA replication and protein sythesis.
• Modification of the activity of numerous enzymes.

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 Actions of insulin on the human metabolism

• It is the main hormone that controls the blood glucose level.

• It stimulates synthesis of glycogen, fat, and protein. (anabolic)
‘’Stimulates glycogenesis in liver and skeletal muscles as well as
lipogenesis in adipose tissue’’
• It inhibits breakdown of glycogen, fat, and protein.
• It increases glucose transport into cells.

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 Insulin for medical purposes
• Insulin is being produced biosynthetically
using recombinant DNA technology.
• More recently, reserchers have succeded
in introducing the human insulin gene into
plants and producing insulin in them. This
technique is set to reduce production
costs.
• Before it was possible to produce insulin
biosynthetically it was aquired from
animals and purified so it could be used as
injections.
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Thanks