HEME


Heme is pronounced as“heem”.
Heme is a derivative of the porphyrin.
Porphyrins are cyclic compounds formed by fusion of 4
pyrrole rings linked by methenyl (=CH-) bridges

Heme is the most important porphyrin containing compound

Heme is a ferroprotoporphyrin Since an atom of iron is present.

Metal ions can bind with nitrogen atoms of pyrrole rings to form complexes.
The pyrrole rings are named as I, II, III, IV and the bridges as alpha, beta, gamma and
delta.

The possible areas of substitution are denoted a 1 to 8
 Structure of Heme

The usual pyrrole rings

substitutions are
Propionyl (–CH2–CH2–
COOH) group

Acetyl (–CH2–COOH)
group
Methyl (–CH3) group
Vinyl (–CH=CH2) group.

Heme Porphyrin ring

STRUCTURE OF HEME
Two additional bonds are on either side of
the plane of the protoporphyrin ring.

5
 Function of Heme

Function: binding O2, electron transfer, oxidation reactions.

Heme is present in:

Hemoglobin
Myoglobin

Cytochromes

Peroxidase
Catalase

Tryptophan pyrrolase
Nitric oxide synthase.
Heme is produced by the combination of iron with a porphyrin ring. Chlorophyll, the
photosynthetic green pigment in plants is magnesium-porphyrin complex.
 HB and MB

Myo - from the Greek word for "muscle“

Hemo - from the Greek word for "blood"
 Myoglobin and hemoglobin are hemeproteins(globular proteins )whose
physiological importance is principally related to their ability to bind
molecular oxygen.
These globins dramatically improve the concentration of molecular
oxygen that can be dissolved in the biological fluids of vertebrates and
some invertebrates.
 Hemoglobin

Hemoglobin is red blood pigment, found exclusively in erythrocytes

It is a conjugated protein (chromoprotein) containing heme as the prosthetic
group & globin as the protein part- apoprotein

Hemoglobin = Heme + globin ( protein )

The basic protein “globin” varies from species to species in
its amino acid composition & sequence, and is responsible for
species-specificity

Polypeptide chains of globin of adult Hb, contain high
content of ‘histidine’ & ‘lysine’ & small amount of isoleucine

Heme containing proteins are characteristic of aerobic organism

The polypeptide chains are usually tetrameric 2à & 2ß chains.

Each gram of Hb contains 3.4 mg of iron.

Normal levels:

Adult male:14 to 16 gm%

Female:13 to 15 gm%
Structure of Hemoglobin
 Hemoglobin

α chain gene is on chromosome 16 beta, gamma and delta
chains on chromosome 11.
Each alpha chain has 141 amino acids.

The beta,gamma and delta chains have 146 amino acids.

36 histidine residues in Hb molecule=for buffering action

58th residue in α chain = distal histidine it is far away from the
iron atom.

87th residue in α chain is = proximal histidine, it lies nearer to
the iron atom
 Iron has 2-oxidation states
COO- COO- COO- COO-
CH2 CH2 CH2 CH2
CH2 CH2 CH2 CH2
H H
C C C C C C
C C C C
H3C C C CH3 H3C C C CH3

C N N C C N N C

HC Fe2+ CH HC Fe3+ CH

C N N C C N N C
H2C H2C
C C C CH3 C C C CH3
C C C C
H C H C
C C C C
H H
CH3 CH=CH2 CH3 CH=CH2

 Ferrohemoglobin (Fe2+)  Ferrihemoglobin (Fe3+), also

 binds O2 known as methemoglobin,
 does not bind O2 11
o When heme is free in solution, it can bind O2.

o But in the presence of a second heme

molecule, O2 will oxidize Fe2+ to Fe3+, so the

first heme no longer binds O2 Fe3+ O

o This process continues until all of the heme
O Fe2+
iron molecules have been oxidized.

o One function of Hb & Hb is to physically

separate the heme molecules,so oxygen doesn’t
oxidize the Fe2+.
O2 binding to Hb

 Tetrametric, 2α-chains & 2β-

 One molecule of Hb can bind with four
molecules of O2.
carries O2 from lungs to tissues
cooperative binding of O2
 required to increase the solubility of O2 in
blood
 Transport of O2 by haemoglobin

Hemoglobin has all the requirements of an ideal
respiratorypigment (Barcroft):
It can transport large quantities of O2 from lungs to the
tissues

Transport of CO2 & protons from tissues to lungs for
excretion
It has great solubility

It can take up and release oxygen at appropriate partial
pressures
It is a powerful buffer.
 Myoglobin (Mb)

It is a single-chain (Monomeric) globular protein of 153 AA

 containing1 heme group

Found in cardiac& skeletal muscle

Mol. wt. 17,000

Serves as O2 reservoir & carrier to increase rate of O2 transport

transports O2 from capillaries to sites of usage (mitochondria)

is found in cytosol within skeletal and heart muscle cells

Structure of Heme
Non-cooperative binding of O2
Myoglobin (Mb)


α-helical content:-this single Polypeptides chain(80%) is folded in to 8 stretches of
α-helices(A-H).
This helix formation is terminated by Proline b/c its 5-membered ring can't be
accommodated in α-helix.

Location of amino acids
Its interior is entirely composed of non polar amino acids W/c are packed &
stabilized by hydrophobic interaction
Its exterior surface from charged amino acids w/c form H-bond to each other and
with water.
 M
b


Binding of Heme


Heme group is found at the crevice of Mb w/c is lined by non

polar AAs except 2 histidine(proximal (F8) his & distal (E7) his).
O2 binding to Mb

Proximal histidine binds directly to heme but distal one not; rather

it helps to stabilize oxygen binding to the Fe 2+.

The globin(protein) portion of Mb provides a microenvironment

for reversible binding of 1 O2 to heme.

 Binding of oxygen to Mb & Hb

Mb can bind only 1O2 molecule b/c it has 1 heme.



Hb can bind 4O2 molecules b/c it has 4 hemes.



The degree of saturation (Y) of these oxygen-binding sites on all Mb or Hb
molecules can vary between zero (all sites are empty) and 100% (all sites are full).

Oxygen dissociation/association curve.

A plot of Y measured at different partial pressures of oxygen (pO2).

The curve is different for Mb & Hb.
 Oxygenbinding?

Oxygen binding is the binding of an oxygen molecule to a specific functional

proteins (Myoglobin & Hemoglobin) for either transport or storage in vivo
 Oxygen Dissociation Curve (ODC)

The binding ability of hemoglobin with oxygen at physiological pO2 (partial

pressure of oxygen) is shown by the oxygen dissociation curve (ODC)
At the oxygen tension in the pulmonary alveoli, the Hb is 97% saturated with
oxygen.
Binding Site Saturation
Myoglobin
100

75

Hemoglobin
50

25

10 20 30 40 50
pO2 (torrs)

The binding of oxygen to myoglobin follows a hyperbolic curve while

the binding of O2 to Hb follows a sigmoidal(S) curve.

Factors affecting oxygen dissociation curve
Heme-heme Interaction & Cooperativity:
The oxygen dissociation curve (ODC) is sigmoid shape.
The binding of O2 to one heme residue increases the affinity
of remaining heme residues for O2.
Thus the affinity of Hb for the last O2 is about 100 times greater than the binding
of the first O2 to Hb.
This is called positive cooperativity=the binding affinity increases as more O2
are bound.
Release of O2 from one heme facilitates the release of O2
from others.
 Effect of 2,3-BPG

Normally the 2,3-bisphosphoglycerate level is 15 +- 1.5 mg/g Hb.

The 2,3-BPG concentration is higher in young children compared to the elderly.
The 2,3-BPG is produced from 1,3-BPG, an intermediate of glycolytic pathway
The 2,3-BPG, preferentially binds to deoxy-Hb and stabilizes the T conformation.
When the T form reverts to the R conformation, the 2,3-BPG is ejected.
During oxygenation, BPG is released
The high oxygen affinity of fetal blood (HbF) is due to the inability of gamma
chains to bind 2,3-BPG.
Adaptation to High Altitude
1. Increase in the number of RBCs
2. Increase in concentration of Hb inside RBCs
3. Increase in BPG
Mechanism of positive cooperativity in Hemoglobin
Binding of O2 to Fe moves proximal Histidine residue and
its attached helix (F)
 Helix F adjusts conformation by movement of a b subunits
(hinge and helix ratchet)
Alters conformation of Iron (Fe) at un-liganded sites.
A ligand (from the Latin ligandum, binding) is a substance (usually a
small molecule), that forms a complex with a biomolecule to serve a
biological purpose.
Allosteric Effects and Cooperativity

Allosteric effects occur when the binding properties of a macromolecule change

as a consequence of a second ligand binding to the macromolecule and altering its
affinity towards the first, or primary, ligand.
2 Types of Allosteric effects
1. Homotropic allosteric effect:. If the two ligands are the same (e.g. 02)
2. Heterotropic allosteric:. If the two ligands are different (e.g. o2
BPG),
Macromolecules that have multiple ligand binding sites
(e.g. Hb), allosteric effects can generate cooperative behavior.
 Allosteric effects are important in the regulation of enzymatic reactions.
 Both allosteric activators (which enhance activity) and allosteric inhibitors
(which reduce activity) are utilized to control enzyme reactions.
 Allosteric effects
Allosteric effects require the presence of two forms of the
macromolecule.
One form, T or tense state, binds the primary ligand
(e.g. oxygen) with low affinity.
The other form, R or relaxed state, binds ligand with
high affinity.
The T and R states are in equilibrium with each other.
In the case of positive cooperativity the fraction of T states
exceeds that of the R state.
The quaternary structure of oxy-Hb is described as R (relaxed) form; & deoxy- Hb is T (tight) form
 Lung surfactant
o It a complex with a unique phospholipid and protein composition.
o Its specific function is to reduce surface tension at the pulmonary air-liquid interface
o Surfactant is composed of 90% lipid and 10% protein molecular weight,
o The most abundant species are phosphatidylcholine, phosphatidylglycerol, and
phosphatidylinositol
o phosphatidylcholine constitutes approximately 70% of the lipid portion of surfactant,
o The lipid content contains primarily phospholipid,
o specifically dipalmitoylphosphatidylcholine (DPPC)
o which is responsible for the biophysical function of surfactant
o This lipid composition is well conserved among vertebrates
 Lecithin(Phosphatidylcholine) Cephalin

Phosphatidylserine
Plasmalogens
 Surfactant
Contain four associated proteins
o surfactant protein (SP)-A, SP-B, SP-C, and SP-D.
o SP-A and SP-D, are hydrophilic and SP-C and B hydrophobic
o  SP-A and SP-D are members of a family of innate immune proteins,
termed collectins
 Surfactant

o Surfactant components are synthesized primarily by:

o The alveolar type II cell, which produces surfactant lipids and
surfactant proteins
o The airway club cell, which synthesizes surfactant proteins SP-A, SP-B,
and SP-D
 Key Functions of Surfactant
1. lowering surface tension at the air–liquid interface and thus
preventing alveolar collapse at end-expiration,
2. Interacting with and subsequent killing of pathogens or preventing
their dissemination, and
3. modulating immune responses
4. Although surfactant is an established treatment for RDS in preterm
infants, no clinical benefit has been shown in adult patients with
ARDS.