BCHET-147

ORGANOMETALLICS,
Indira Gandhi National BIOINORGANIC CHEMISTRY,
Open University
School of Sciences POLYNUCLEAR HYDROCARBONS
AND UV, IR SPECTROSCOPY

Block

2
BIOINORGANIC CHEMISTRY

UNIT 5
Metals in Biological System 73

UNIT 6
Function of s-block Metals 89

UNIT 7
Role of Iron in Oxygen Transport 97
Course Design Committee

Dr Pritam Mukhopadhyay School of Sciences,

School of Physical Sciences, IGNOU, New Delhi 110068
Jawaharlal Nehru University,
New Delhi Prof. M.S. Nathawat
Prof. Sunita Malhotra
Dr. Preeti Malhotra
Department of Chemistry, Prof. Bharat Inder Fozdar
University of Delhi, Delhi Prof. Javed A. Farooqi
Prof. Sanjiv Kumar
Dr. Sushmita Choudhury (Retd.)
Prof. Lalita S. Kumar
Gargi College, University of Delhi,
New Delhi Prof. Kamalika Banerjee

Dr. Sangeeta Pandita

Department of Chemistry,
Zakir Husain Delhi College,
University of Delhi, Delhi

Block Preparation Team

Prof. Kamalika Banerjee Dr. Sushmita Choudhury (Retd.), (Editor)
School of Sciences, IGNOU Gargi College, University of Delhi,
New Delhi

Course Coordinator: Prof. Kamalika Banerjee

Production
Mr. Rajiv Girdher Mr. Hemant Kumar
AR (P), MPDD, IGNOU SO(P), MPDD, IGNOU

Acknowledgements: Sh. Sarabjeet Singh and Sh. Deepak Kumar for word processing and Sh.
Sarabjeet Singh for CRC preparation.

July, 2022
 Indira Gandhi National Open University, 2022

ISBN: 978-93-5568-436-3

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 BLOCK-2: BIOINORGANIC CHEMISTRY

This is the second block of this course and here you will be introduced to bioinorganic
chemistry which is of immense interest as it involves living systems. This block has three
units.

Unit 5 deals with bioinorganic chemistry followed by metals and non metals in biological
system. The essential, non essential as well as trace and toxic metals have been discussed.

In unit 6 the sodium-potassium pump and the role of magnesium ions in chlorophyll have
been dealt with.

In the last unit of this block, that is unit 7, the role of iron in oxygen transport along with
details of hemoglobin and myoglobin have been discussed.

Expected Learning Outcomes

After studying this block, you will be able to

 understand the importance of bioinorganic chemistry;

 appreciate the presence of metals & non-metals in biological system;

 define the essential, non-essential, trace & toxic metals;

 describe the Na+-K+ Pump;

 understand the role of magnesium ions in chlorophyll;

 describe the salient features of hemoglobin; and

 discuss the special characteristic of myoglobin alongwith its structure.

Unit 5 Metals in Biological System

UNIT 5

METALS IN BIOLOGICAL
SYSTEM

Structure
5.1 Introduction Essential, Non essential

Expected Learning Outcomes Trace & Toxic Metals

5.2 Bio-Inorganic Chemistry 5.4 Summary

5.3 Metals & Non-metals in 5.5 Terminal Questions

Biological System
5.6 Answers

5.1 INTRODUCTION
The last block had four units where you learnt about the important inorganic
reagents, organometallic compounds alongwith their nomenclature and the
metal carbonyl compounds. The applications of inorganic chemistry to living
systems is one of the most rapidly developing areas in modern inorganic
chemistry. There are vital “inorganic” links in the processes that are discussed
in this block on bioinorganic chemistry. Of course, this interdisciplinary subject
is still in its developmental stage and is very futuristic in nature. Scientists are
still trying to find out the answers to many unsolved puzzles of the living
system. In this unit you will be studying about metals in biological system in
details.

Expected Learning Outcomes

After studying this unit you should be able to:

 understand the importance of bioinorganic chemistry;

 appreciate the presence of metals & non-metals in biological system; and

 define the essential, non-essential, trace & toxic metals.

73
Block 2 Bio-Inorganic Chemistry

5.2 BIOINORGANIC CHEMISTRY

Bioinorganic chemistry involves the interface of inorganic chemistry and
biology. The term “inorganic biochemistry” or “bioinorganic chemistry” is used
in the latter part of the twentieth century which includes studies on the
biological functions of typically “inorganic elements”,  the metals in particular.
Many inorganic elements are essential to life processes. Different elements
are taken up selectively by different cells and other components inside the
cells, and utilized. The basics of coordination chemistry have been discussed
in Units 4 and 5 of BCHCT 137 which you may recapitulate before reading this
unit. Here you will be discussing the functions of the metals in the biological
environment in context with the coordination chemistry which you may have
learnt earlier. The mystery of living systems have always attracted scientists
who tried to unravel it with the help of experimental research. In this process,
model compounds with structures almost the same as in living systems have
been tried to find the mechanism of reactions in natural systems. The term
“bioinorganic” obviously has lot of contradictions. The source of this are the
misconceptions that have been there when modern science evolved. In the
early 19th century, chemistry was still divided into “organic” chemistry which
comprised of substances which were found in “organisms”, and an “inorganic”
chemistry which comprised of “dead matter”. But then in 1828, synthesis of
“organic” urea from “inorganic” ammonium cyanate (NH4OCN) changed the
basis of this terminology.

By 1960, bioinorganic Sophisticated and advanced analytical techniques have revealed that apart
chemistry became an from carbon many metals and non-metals are present in biochemical
independent and processes and they are important for life. Even trace elements could be
highly interdisciplinary detected in these processes. Thus we can say that bioinorganic chemistry is
research area.
mainly the study of the function of inorganic substances in living systems,
alongwith the transport, speciation and thereby, mineralization of inorganic
materials and their use in mechanical therapy and diagnosis.

The major areas of study of bioinorganic chemistry may be broadly identified

as :

(i) biological functions of different elements which are mostly known to form
inorganic compounds;

(ii) to find out if these elements and their compounds are toxic and how to
overcome the toxic effects;

(iii) metal ion transport and storage in living systems;

(iv) use of metals as probes and drugs;

(v) artificial fixation of nitrogen and other applications.

The following two chemical processes involved in the chemistry of life in which
metal ions play an active role are:

(i) Using solar energy to facilitate chemical reactions that produce oxygen
and reduced organic compounds from carbon dioxide and water
74 (photosynthesis)
Unit 5 Metals in Biological System

(ii) Oxidation of organic matter to produce carbon dioxide, water and

energy (respiration).

The “reaction flask” of living organisms is the cell. Each cell is characterized by
DNA is deoxyribose
an outer membrane whose function is to contain a highly organized chemical nucleic acid RNA is
system and to monitor the influx of needed reagents. Within the cell ribose nucleic acid.
membrane are several organelles immersed in cellular fluid, cytoplasm. Two of ATP is adenosine
these organelles, the nucleus and mitochondria, are the focus of most of the triphosphate.
chemistry. The nucleus is surrounded by a nuclear membrane, within which
occur the process concerned with DNA-DNA replication, RNA synthesis and
membrane synthesis. The other important organelles are the mitochondria
within which occur the redox/electron transfer processes important in the
combustion of glucose and synthesis of ATP. In animals the mitochondria are
the sole centres of energy generation in the cell. The cells of green plants
contain in addition chloroplasts which contain chlorophyll. This chlorophyll only
enables the light sensitized phosphorylation reaction associated with ATP
generation.

In the dark, the mitochondria of plant cells maintain this regeneration though at
a lower rate than in animals. This is because in green plant cells it is rarely
found. The other organelles are lysosomes and golgi bodies involved in
digestion and excretion respectively. The endoplasmic reticulum is an
intracellular network of channels for transport of proteins synthesised by
ribosomes that surround the surface of the channels.

An important feature of living systems is their unique dependence upon kinetic

stability for their existence. Organisms must maintain steady states far from
equilibrium by a continuous flow of nutrients and energy in a variable
environment. All are thermodynamically unstable. They would burn up
immediately to carbon dioxide and water if the system came to thermodynamic
equilibrium. Thus to a living organism reaching equilibrium is equivalent to
death. Life processes depend upon the ability to restrict these thermodynamic
tendencies and allow kinetics to control and produce energy as needed. So,
apart from capturing solar energy, it is important to use catalysts to release
that energy in controlled manner. Examples of such catalysts are enzymes
which control the synthesis and degradation of biologically important
molecules. Metal ions play a crucial role for the activity of those enzymes.
Metal containing compounds are also important in the process of chemical and
energy transfer, reactions involving transport of oxygen to the site of oxidation
and various redox reactions. Most of the reactions for obtaining energy for
living systems are basically inorganic. The reactions are mediated and made
possible by complex biochemical processes.

Inorganic elements can also be artificially introduced to the living system in

diagnostic techniques like barium meal X-ray, MRI (Magnetic resonance
imaging, suitable for paramagnetic metal ion) and as medicines e.g., platinum
complexes in oncology, gold complexes in arthiritis and lithium salts as
antidepressants. In these ways, bioinorganic chemistry provides many
interesting insights about the functions of inorganic substances in living
systems. In the next section you are going to study about the metals and non-
metals in biological system.
75
Block 2 Bio-Inorganic Chemistry

Before moving to the next section, please try to solve the following SAQ.

SAQ 1
Give any two major areas of study of bioinorganic chemistry.

5.3 METALS & NON-METALS IN BIOLOGICAL

SYSTEM
In this section you are going to study the metals and non-metals in biological
system.

5.3.1 Essential, Non-essential Elements

Now let us discuss the essential elements in biological systems.

Essential elements are those elements which have a specific role

indispensable to sustain life in a living organism. The normal functioning of the
living system will suffer if there is absence or even deficiency of the element.

About thirty elements appear essential to some form of life (Table 5.1). The
most abundant biological element is hydrogen which constitutes 63% of the
atoms in a human body. Next comes oxygen (25 .5%), carbon (9.5%) and
nitrogen (1.4%). The following seven elements in order of their abundance in
humans are Ca, P, Na, K, S, Cl and Mg; together they constitute about 0.6%.

Table 5.1: Essential elements in living organisms

bulk elements : ; others are trace elements

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

H B C N O F

Na Mg Si P S Cl

K Ca V Cr Mn Fe Co Ni Cu Zn As Se Br

Mo Sn I

There are nearly 40 elements in the periodic table which are biologically
active. Any species responding to or involved in a biological system is termed
biologically active. The heaviest biologically active metals and non-metals are
molybdenum and iodine whilst the lightest counterparts are lithium and
hydrogen. The following elements are essential of human life.

Metals: Na, Ca, Mg, V, Cr, Mn, Fe, Co, Cu, Zn, Mo, Sn.
Non metals: H, C, N, O, P, S, Se, F, Cl, Br, I.

The 11 most abundant elements are H, O, C, N, Na, K, Ca, Mg, P, S, Cl; the
next seven are less abundant - Mn, Fe, Co, Cu, Zn, Mo and I whilst the
remainder occur in ultratraces. The amount of these elements present in a
70 kg man are shown in Table 5.2 below.
76
Unit 5 Metals in Biological System

Table 5.2: Amounts of essential elements present in a 70 kg man

Oxygen - 45.5 kg Ca – 1050 g
Carbon – 12.6 kg K – 140 g
Hydrogen – 7 kg Na – 105 g
Nitrogen – 2.1 kg Mg – 35 g
Phosphorus – 700 g Fe – 4.2 g
Zn – 2.3
Mn, Cr, Mo, Co, Cu < 1g
Eg. Cu – 0.11 g
Mn – 0.02 g

Hydrogen, oxygen and carbon are present in water, carbohydrates, proteins

etc. Nitrogen is an essential constituent of proteins. Phosphorus is present in
ATP, ADP, DNA, RNA, bones, teeth etc. Sulphur is present in some amino
acids and vitamins. Sodium, potassium, magnesium, chlorine are present in
body fluids and cytoplasm. Sodium ion (Na+) is the major component of
extracellular fluid and potassium (K+) of intracellular fluid. Together they act as
charge carriers and are involved in osmotic balance. That is, they maintain a
constant osmotic pressure on either side of cell membrane. The acid-base
equilibrium is maintained by them. They also control sensitivity of nerves and
muscles along with calcium ions (Ca2+) and magnesium ions (Mg2+). Calcium
is required for bones and teeth and maintain correct rhythm of heart beat and
converts fibrinogen to fibrin. It also stabilises protein structure. Mg2+ are found
complexed with nucleic acids and are necessary for nerve-impulse
transmission. All the phosphate transfer enzymes need Mg2+. It is also an
essential constituent of chlorophyll.

Manganese (Mn) and cobalt (Co) are present in enzymes, for example,
arginase involved degradation of protein. Co is present in Vitamin B12. Copper
(Cu) is a constituent of metalloenzymes involved in electron transfer. Iron (Fe)
is present in hemoglobin, myoglobin, cytochromes which act as O2 carrier, O2
storehouse and electron transfer respectively. Zinc (Zn) is present in enzymes
particularly hydrolases and in insulin. Molybdenum (Mo) helps in nitrogen
fixation in plants while iodine (I) is involved for proper functioning of thyroid
gland.

We may classify the elements in a way that suggests their function in the
complex dynamic system of a living cell. Let us now discuss a system that
classifies elements according to their occurrence in three different biological
environments.

 Firstly, it may be in the extracullar fluids and outer wall of cell-membrane –

Na, Ca, Cu, Mo, Cl, Al.
 Secondly, it may be inside the cell, starting from inner wall of cell
membrane in the aqueous phase known as cytoplasmic fluid - K, Mg, Co,
Zn, P, S
 Also it may be present organelles inside the cell – K, Mg, Fe, Co, Zn, Mn,
P, S

However, this classification is not very rigid.

77
Block 2 Bio-Inorganic Chemistry

Next, let us dscuss the classification of the elements on the basis of

mechanism of action as:
 Essential elements: they are absolutely necessary for life processes and
are needed in large quantites e.g. O, C, H, N, P, Na, K, Mg, Cl, Ca, S
 Trace elements: these are essential but are present in trace amounts - I,
Fe, Cu, Zn, Co, Mn, Mo.
 Non-essential: they may be present in biosystem, but in their absence
other elements may serve the same purpose e.g. Al, Ba, Sn, Sr.
 Toxic: they disturb the normal function of biological system e.g. Hg, Pb,
Cd, As etc.

Next let us understand what are biomolecules. Biomolecules containing one

or more metallic elements are called metallo biomolecules. These are natural
products and usually complex coordination compounds, whose active sites
are involved in different processes e.g., electron transfer, O2 transfer,
catalysis etc. Most biochemical processes are related to the chemistry of
hydrogen and elements of the second and third periods of the periodic table.
Every cell (animal, plant, microbial) contain the same elements in the same
proportion. Six nonmetals (H, C, N, O, P, S) provide the structural elements to
the protoplasm. The relative geochemical abundance and intrinsic chemical
properties like, (i) small atomic radius, (ii) catenation, (iii) ability to from
multiple bonds of these six elements suggest their unique suitability for
building blocks of life. Table 5.3 gives an idea of the important
metallobiomolecules

Table 5.3: Some representative examples of different metallobimolecules

METALLOBIOMOLECULES

METALLOPROTEINS NON-PROTEINS

Metal storage and transfer

Transport and Storage Metalloenzymes
Siderophores (Fe)

Photoredox
Electron transport Hydrolases
Chlorophyll (Mg)
Cytochromes (Fe) Phosphates (Mg, Zn, Cu)
Iron-sulphur (Fe) Aminopeptidases (Mg, Zn)
Copper blue (Cu) Carboxypeptidases (Zn)
Dioxygen management Oxidoreductases
Haemoglobin (Fe) Oxidases (Fe, Cu)
Myoglobin (Fe) Reductases (Fe, Cu, Mo)
Haemerythrin (Fe) Superoxide dismutases (Fe, Cu, Mn)
Haemocyanin (Cu) Isomerases and synthetases
Metal management Vitamin B12 coenzyme (Co)
Ferritin (Fe)
Transferases
Transferrin (Fe)
Kinases (Mg)
Ceruloplasmin (Cu)
Lyases
Enolases (Mg)
Ligases
Glutamine synthetase (Mg, Mn)
78
Unit 5 Metals in Biological System

The number of elements that are biologically important comprises a relative

small fraction of the known elements. Geochemical abundance is an important
parameter. All essential elements are abundant in the earth’s crust. Four
elements, Al, Si, Ti, Zr are abundant but not essential because at biological pH
they form insoluble oxides. They also form very weak complexes with
biological species. Natural abundance limits availability of elements for use in
biosystem. The metals for importance in enzymes are mainly those of the first
transition series and other elements of importance are relatively the light ones.
Natural abundance of elements decreases with increase in atomic number.
The heavier elements are not very important as far as biological activity is
concerned. The conclusion is inescapable “Life evolved utilizing elements that
are abundant and available to it and became dependent upon them”. The rarer
elements were not used by living systems because they were not available;
neither did the system evolve mechanisms to cope with them. They were later
introduced to the environment and ecosystem by human activities.

Most of essential metals except Na, K cannot occur in the biosystem in the
free state as under the prevailing pH conditions they get hydrolysed and
precipitated. They must be present and transported as complexes. Most of the
essential and trace metals are transition metals which are capable of forming
complexes and chelates with species like citrate, tartrate, lactate, amino acids
present in the plasma. Many metal complexes of biological macromolecules
have been isolated in the free state for example hemoglobin, myoglobin,
cyanocobalamine, chlorophyll a, cytochrome-c etc. The presence of others or
their formation as intermediate products taking part in metabolic processes of
living system has been inferred from a catalytic study of enzymes.

A successful organism must have a subtle command over a complex pattern

of reactions that proceed at high but controlled rates. Loss of this control
leads to the organism’s disease and death. Life depends on how these
reactions are controlled and thus the role of catalysts is important. These
catalysts are enzymes which are polypeptides i.e. macromolecules, formed

from -amino acids linked by a peptide bond . The term “peptide

residue” denotes that part of the amino acid that remains in the chain once
the peptide link is formed by elimination of H2O. Enzymes are named
according to the reactions they catalyze e.g. hydrolase, oxidase, reductase
etc.

The naturally occurring amino acids have different side chains eg. alkyls,-
COOH, -NH2, -OH, -SH. These functional groups confer hydrophobic and
hydrophilic character, Bronsted acidity/basicity, Lewis acidity to complex with
a metal. About 30% enzymes are metalloenzymes having a metal atom as the
active site. The functional groups modify the immediate environment. The
protein part of the enzyme is called apoenzyme and the non-protein part, the
prosthetic group. Enzymes not only control the rate of a reaction but by
favouring certain geometries in the Transition State (TS) can lower the
activation energy to favour the formation of the desired product. Metal ions
also occur in transport and storage proteins and in non-protein complexes.

Life originally appeared on earth in an overall reducing atmosphere. As shown

by Miller and Urey (1953), lightning discharges through a mixture of methane,
water, ammonia and hydrogen to produce significant amounts of amino acids 79
Block 2 Bio-Inorganic Chemistry

essential to life processes. Abelson and others later showed (1960s) that
solar ultra violet radiation not filtered by the earth's ozonosphere acts on a
mixture of carbon monoxide, carbondioxide, nitrogen and a small amount of
hydrogen producing, among others, hydrogen cyanide. This molecule can
subsequently give rise to glycine, adenine, and other biologically significant
molecules. These compounds, formed in the primitive earth, might have
undergone a series of complex chemical evolution over millions of years to
produce living organisms which could replicate themselves and undergo
biological evolutions (nearly 3.5 billion years ago). Photosynthetic organisms
appeared approximately 2.5 billion years ago when the atmosphere became
gradually rich in oxygen and ultimately sustained aerobic forms of life. The
diverse biological world of today has largely developed with this form of life
and is a unique example of nature's delicate balance between constructive
and destructive oxidation by oxygen. The amazingly complex and yet
precisely synchronized biological processes involve, in addition to H, C, N, O,
and P, other elements abundant on the earth, particularly a large number of
metals, in both bulk and in trace. As more and more finer details regarding
these roles get unveiled, the emphasis on a particular area of study generates
new terminologies to justify the thrust area.

5.3.2 Trace & Toxic Metals

The living system also requires small amounts of other elements as shown in
Table 5.2. The requirement of these elements may be very low (i.e., 104 g
mole or less in a 75 kg adult) or moderate (e.g., 101 g mole). All these
elements are classed as essential trace elements of which in top priority; iron,
copper and zinc. Elements present in exceedingly small amounts are called
ultra-trace elements, for example, Mn, Mo, Co, Ni and V.

Hydrogen are involved in the biological system and the s-block elements
sodium, magnesium, copper and calcium. Na, K, Ca and Mg, are the most
abundant metal ions in living systems. They occur at fairly high concentration
in most cells and constitute 99% of the metal content (more than 1% of the
body weight) in man.

Hydrogen

It is the most abundant element in the biosphere. The small atoms can
effectively “seal” the residual valences of carbon, nitrogen, and oxygen without
any structural strain. When bonded to nitrogen and oxygen, hydrogen can
form hydrogen bond; these weak but extensive bonds are crucial in stabilizing
many biological systems, for example, the double helix of DNA. The weakness
of the bonds is an advantage in such systems since the helix must unwind
during replication. The life supporting properties of water and many other
functions like operations of several enzyme are also dependent on hydrogen
bond.

Sodium and Potassium

In spite of their established chemical similarity, the elements differ in their

ability to penetrate cell membranes presumably due to their different ionic size.
80 The metals also differ in their transport mechanism and efficiency to activate
Unit 5 Metals in Biological System

enzymes. Sodium ions are more abundant outside the cells, for example in the
blood plasma and in the extracellular fluid surrounding the cells. They
participate in transmitting nerve signals and regulate the flow of water across
cell membranes. The transport of sugars and amino acids into cells is also
influenced by sodium ions (Na+).

On the other hand, potassium ions occur at a higher concentration inside cells.
They also activate many enzymes and participate in the oxidation of glucose
to produce ATP. Both sodium and potassium ions are also involved in the
transmission of nerve signals.

Magnesium

The involvement of magnesium in photosynthesis by plants is well-known.

Magnesium is also an essential element for animal physiology. The major
portion of magnesium is an adult human is in the skeleton while the rest is
inside the cells. The Mg2+ ion in cells is the next most important cation after K+.
The small doubly charged cation has a high charge/size ratio and is strongly
hydrated - mostly to [Mg(H2O)6]2+ which binds the ROPO3H – groups in
nucleotides (ATP, ADP) and polynucleotides (DNA, RNA). The ion may form
strong complexes with oxygens on various phosphate groups; such complexes
appear to be important in storing energy and use of the same when
necessary. All enzymes utilizing ATP in phosphate transfer require Mg2+ as
cofactor.

Intracellular magnesium ions are also essential to keep ribosomes intact.

Ribosomes are RNA-protein complexes of particle size of about 20,000 pm
diameter which occur in the cytoplasm and function as sites for protein During the period of
synthesis. growth, deposition of
bone must exceed
Calcium resorption. With
ageing, resorption
It is the major constituent of bones and teeth. It is also important in predominates and
neuromuscular function, interneuronal transmission, maintenance of cell causes loss of about
membrane integrity and blood coagulation. The calcium present in bones gets 15-30 mg per day.
continuously solubilized and redeposited in our body to the extent of about 400 Vitamin D promotes
mg per day. The concentration of calcium ions (Ca2+) in plasma is closely deposition of bone.
maintained around 100 mg/L by means of two hormones namely, calcitonin This vitamin is
synthesized by the
and parathyroid hormone. Together they help in building and mobilising bone
action of ultra violet
structure. light on 7-
dehydrocholesterol.
Calcium ions in the cytoplasm of muscle fibres play an important regulatory
So inadequate
role in muscle functioning. Clotting of blood is also influenced by the presence exposure to sunlight
of Ca2+ ions. Blood contains prothrombin, a soluble protein. This is converted may result in
to the enzyme thrombin by the action of prothrombin activator in the presence diseases of the bone
of Ca2+ ion. Thrombin, again aided by Ca2+, now clots the blood by converting like ricket in childhood
its soluble fibrinogen into insoluble fibrin as shown below: and softening of
bones (osteomalacia)
Prothrombin activator in later life.
Prothrombin Thrombin
+ Ca2+
+ Ca2+

Fibrinogen Fibrin
81
Block 2 Bio-Inorganic Chemistry

It is interesting to note that the distribution of essential elements in the

biosphere is not in harmony with the abundance of the elements in the earth‘s
crust. About 88% of the earth‘s crust is made from the four most abundant
elements oxygen, silicon, aluminium and iron. In living cells, more than 98% is
derived from the four elements hydrogen, carbon, nitrogen and oxygen. On the
other hand, the concentrations of different ions in sea-water show a similarities
with those in living systems, for example, the blood plasma. Thus life probably
originated in the sea and evolved thereafter. The process of evolution
continued with the relatively abundant elements which were found suitable for
sustained development of the process. The metal ions chosen were able to
form thermodynamically stable units in the active centres of proteins
(metalloproteins) or other biomolecules. At the same time, they had to be
kinetically labile, be able to assemble and disassemble very fast under
physiological conditions. Titanium and zirconium, in spite of their high
abundance, are not found in living cells as the metals are readily precipitated
as insoluble hydrated oxides at our body pH. Also the metals have a uniform
high +4 oxidation state unsuitable for reversible electron transfer and other
reversible biological processes.
The health response of an organism to an essential element follows the
general type of curve shown in Fig. 5.1; details of the curve vary from one
element to another. At and around zero intake of the element, the condition
may be anything between bare survival to death. As the dose is slowly
increased, the organism gradually reaches a stage of smooth functioning. But
an excessive dose will gradually result in toxicity by the same element and
ultimately lead to death. For some elements, the safe dose and the toxic dose
may be quite close, for example 50 to 200 g for selenium (Table 5.4).

Fig. 5.2: Nature of variation of response to increasing dose of an essential

element.

Table 5.4: Recommended daily doses of some elements for an adult

human being (~70 kg)

Element Intake (per day) Element Intake (per day)

Iron 10 mg (male) Fluorine 1.5-4.0 mg
18 mg (female) Molybdenum 150-500 g
Zinc 15 mg Iodine 150 g
Manganese 2.5-5.0 mg Chromium 50-200 g

82 Copper 2.0-3.0 mg Selenium 50-200 g

Unit 5 Metals in Biological System

Deficiency or excess of these elements may give rise to metabolic disorders,

inhibited growth and other diseases.

Sometimes one element strongly interferes with the uptake of another element
an this is known as antagonism. The copper containing enzyme monoamine
oxidase is involved in the cross-linking of polypeptide chains responsible for
structural strength of arteries. As a consequence, copper deficiency in
herbivorous animals causes anemia, loss of pigmentation and ultimately death
owing to arterial lesions. However, even when the animals are supplied with
pasture having a plentiful supply of copper, the take may be still hampered if
the soil contains a high level of molybdenum – presumably owing to the
formation of highly insoluble copper thiomolybdate in the gut.

Let us look into specific examples of toxicity of metal ion in biochemical

system. Metal ions are essential for life processes only in limited quantity. The
concentration of the essential metal ions inside the cell is controlled by
enzyme regulated transport mechanisms and excess unused metal ion is
ejected. The biosystem does not have transport mechanism for some
(impurity) elements and these cause disorder. Such metal ions are toxic, the
highly toxic ones are generally heavy metals of second and third transition
series (soft acids) and metalloids like arsenic. Toxicity is of two types, namely
chronic toxicity and acute toxicity. Chronic toxicity arises due to high rate of
turnover of metal ion in the body. Whenever metal ions are in excess in the
body, they are stored in specific sites to a limited extent and the excess metal
ion interferes in body processes. Acute toxicity arises due to certain chemical
activity like deactivation of enzymes and other biomolecules. Let us
understand this in detail now.

(i) The metal may get bound to coordinating site of the enzymes, which are
binding sites of essential metal ions. Thereafter the essential biological
function of the enzyme is lost resulting in its deactivation

} L
2+ +
} LH + M M + 2H L = O, N, S
} LH }L
Enzyme

(ii) Deactivation of an enzyme can be due to displacement of metal ion

present in the enzyme, which is essential for its catalytic activity by a
toxic metal ion e.g. replacement of Zn2+ by Pb2+, Cd2+, Hg2+.

} L } L
n+ n+ n+ n+
M + M' M' + M
} L } L

As the external metal ion is not biochemically active, the activity of the
enzyme is lost. Common example is Cd2+ replacing Zn2+.

(iii) There may be cases when the external metal ion may change
conformation of biomolecule and hence its biological activity.

(iv) The metal ion can get bound to DNA causing a change in base
sequence, leading to transmission of wrong genetic information from 83
Block 2 Bio-Inorganic Chemistry

DNA which produces faulty protein and enzymes. This may give rise to
birth defects too.

(v) The metal ion can bind to DNA stimulating its replication resulting in
uncontrolled cell growth i.e., formation of malignant tumours.

Lead

This enters the biosystem through food, water and air. Lead pollution can be
caused by factories, particularly those manufacturing batteries. The antiknock
compound Pb(Et)4, namely tetraethyl lead used earlier in gasoline was present
in exhaust gases of automobiles and was a source of lead (Pb) in the
atmosphere. Lead is also present in paints and pigments. Lead gets deposited
in the softer tissues and passes to the blood stream. Pb2+ has strong affinity
for the –SH group of enzymes and hence gets bound to these and deactivates
them. The principal indicators of lead level in human beings is lead in blood
expressed as g/mL or mol/L. At concentration as low as 10 g/mL it inhibits
some enzymes when the concentration becomes higher, it affects digestive,
muscular, nervous and other organs. Lead affects the biosynthesis of bones
as Pb2+ replaces Ca2+ in bone. Deposition of lead in brain results to
irreversible brain damage. Lead also damages kidney, liver and intestinal tract.

The major biochemical effect has been attributed to its influence with heme
synthesis giving rise to hematological damage. It lowers the formation of
-aminolevulinic acid (ALA) and its conversion to porphobilinogen is inhibited.
Lead deactivates the enzyme ALA dehydrase and overall effect is disruption of
synthesis of hemoglobin, other respiratory pigments and cyctochromes.
Thereby deficiency of hemoglobin leads to anemia. Methods for removal of
lead from the body depend on formation of coordination compounds which are
excreted. Common antidotes are calcium sodium salt of EDTA (CaNa2EDTA),
succimer (Dimethyl succinic acid or DMSA), 2,3-dimercapto-1-propanol
(British anti-Lewisite (BAL) and d-penicillamine. The probable coordination
compounds of lead ions with some antidotes are shown below;

COO
H2C CHCH2OH
HC S
S S Pb
Pb with BAL H3C C S with d-penicillanine
S S CH3
H 2C CHCH2OH

Arsenic

Arsenic (As) poisoning is caused through air and water (mainly ground water)
pollution. Elemental arsenic is not absorbed through the intenstinal wall and is
not toxic. As(III) is more toxic then As(V). Poisoning causes gastroenteritis,
skin lesion and skin cancer. As(III) shows toxicity due to binding with the SH
group of the enzyme pyruvate dehydrogenase, which is essential for ATP
synthesis. At higher concentration. As(III) causes coagulation of proteins
probably by binding to SH group of the proteins. It also binds to the protein
84 keratin in the hair affecting its texture.
Unit 5 Metals in Biological System

Excess arsenic affects the biosynthesis of ATP in another way. It is chemically

similar to phosphorus and interferes in stepwise synthesis of ATP. In presence
of AsO33, phosphorylation of glyceraldehyde-3-phosphate to give 1,3-
disphosphoglycerate does not occur, rather 1-arseno-3-phosphoglycerate is
formed and further oxidative phosphorylation leading to ATP formation does
not occur. The main biochemical actions of arsenic are as follows:

 deactivation of enzymes e.g. pyruvate dehydrohenase system, citric acid

cycle is also inhibited apart from others; DMSA is dimethyl
succinic acid.
 coagulation of protein; and
 uncoupling of phosphorylation.

A lethal dose of the more toxic form of arsenic is 125 mg/kg body weight but
for less toxic forms it is upto 200 mg/kg body weight. The major antidotes for
arsenic poisoning include chemicals having SH groups which can bind to
arsenic forming chelate e.g. (British anti-Lewisite (BAL), succimer (DMSA).

Cadmium

Cadmium (Cd) is a very toxic metal with a lethal dose of greater than 350 mg
at a time. It enters the body system through the food chain (40 mg/day) and
industrial fumes. Tobacco contains nearly 1g Cd/g. Thus nearly 2-4 g of Cd
are inhaled per packet of cigarette. This is the major source of inhaled Cd. 25-
50% of inhaled Cd is absorbed into the body while the absorption from food is
almost 6%. Other sources of Cd are from plating, pigment and battery
manufacturing industries. Inhaling Cd laden dust leads to respiratory problem.
Cd accumulates in liver and kidneys and may produce irreversible damage.
Certain compounds of Cd are carcinogenic. The main disease associated with
Cd poisoning is Itai-Itai disease which was observed in the Jinzu river basin
(Japan). This water was used for irrigation and patients developed this disease
after eating rice grown on fields irrigated with this water. The disease is
associated with brittle or hollow bones, high rate of fracture, osteoporosis and
intense bone-associated pain. It is very interesting to note that Cd cannot pass
through the placental membrane and thus cannot harm new born babies.
Excess Cd can displace Zn from enzymes and deactivate them causing
metabolic disorders. Regular intake of Zn provides protection against Cd
poisoning.

The average amount of Cd accumulated in the body over a life time is nearly
30 mg. Metallothioneins (MTs) are low molecular weight cadmium binding
proteins present in animal and human tissues. The deposited Cd is taken up
by MTs and Cd binds to its SH group thereby getting trapped and its toxicity
controlled. The protein bound Cd gets accumulated in the kidneys and
damages them. Long-term cadmium exposure gives rise to kidney or bone
disease, reproductive toxicity and cancer in animals and human. Its antidotes
are (British anti-Lewisite (BAL) and CaNa2EDTA.

Mercury

Mercury (Hg) is the most toxic heavy metal and may enter the biological
system as Hg vapour, mercury salts or alkyl/aryl mercury compounds. The
major source is industrial processes involving mercury compounds and 85
Block 2 Bio-Inorganic Chemistry

organomercurials used as fungicides. Upto 80% of Hg may be absorbed

depending on its chemical form. The methyl mercury compounds are most
extensively absorbed while for inorganic mercury compounds the absorption is
nearly 15% mercury in blood and hair can be used for estimating the body
burden. A blood Hg level of 20 g/100 mL corresponds to a daily consumption
of 200-300 g of Hg. The distribution of mercury in different organs follow the
order liver, kidney, brain, heart, lungs. Its presence is least in the muscles.

The toxicity of Hg depends on its chemical form. In the liquid form it is not
absorbed by the human systems and is not toxic. However Hg vapours is
absorbed into the blood and enter the brain. Hg(II) is toxic but cannot enter the
cell as it cannot pass through the membrane. Hg(I) forms and insoluble
chloride with gastric juice in stomach and is not highly toxic. However Hg(I)
can enter the cell and get oxidized to Hg(II) and show high toxicity. Hg(II) like
Cd(II) and Pb(II) has affinity for SH group of enzymes as well as proteins and
can deactivate them. It also inhibits the synthesis of  aminolevulinic acid by
deactivating specific enzymes and thereby decreases synthesis of heme. It
can also form bonds with heme and serum albumin. The most toxic species
are the organomercurials headed by methyl mercury (CH3Hg+). Mercury salts
are converted into methylmercury in the biochemical system by the action of
Vit B12. The CoC bond in Vit B12 is weak and can be readily transferred to the
substrate causing alkylation. This methylmercury is taken up by aquatic plants
and fish and finally enters animals and human bodies. The higher toxicity of
organomercurials is due to their nonpolar nature, this makes them fat soluble
and they readily pass through the biological membrane and spread all over the
body. The HgC bond does not break easily hence CH3Hg+ is not excreted
and retained in kidneys, brain, heart and muscles. CH3Hg+ can penetrate
through the placenta and get accumulated in foetal tissue. Methylmercury gets
attached to the cell membrane and hence the transport of sugar through the
membrane is lowered whereas external passage of potassium ions (K+) is
increased. Thus the brain cells do not get sufficient energy and K+controlled
nerve impulse transmission is impaired leading to tumours and mental
instability. Such effects are observed distinctly when the concentration of
mercury in the brain exceeds 20 g/g. The effect is more serious in babies. It
may cause irreversible damage to central nervous system is irreversible
leading to cerebral palsy, mental retardation and convulsions. CH3Hg+
poisoning also gives rise to segregation of chromosomes, chromosome
breakage in cells and inhibited cell division.

The toxic effect of mercury was first realized when 121 persons in Japan were
affected by a disease causing tremors and mental imbalance. They were
residents near the Minamata bay where the effluents of the factory containing
mercury compounds and CH3Hg+ was discharged. Though the concentration
of CH3Hg+ was low it got accumulated in significant concentration in fish.
People eating the fish were seriously affected and many babies were born with
cerebral defects and this was referred as Minamata disaster.

So in this section you have learnt all about the metals and non metals in
biological system. Before moving to the next section, please try to solve the
following SAQ.
86
Unit 5 Metals in Biological System

SAQ 2
What are the roles of sodium and potassium metals in biological system?

5.4 SUMMARY
In this unit, we first discussed the importance of bioinorganic chemistry. Then
the different metals and non-metals in biological system along with the
essential, non-essential, trace & toxic metals have been taken up. These will
help you to understand the role of various metals in the biological system. In
the next unit we will talk about the functions of s-block metals.

5.5 TERMINAL QUESTIONS

1. What is bioinorganic chemistry?

2. What is the role of manganese and cobalt in biological system?

3. Which metal ion has role in blood clotting and how does it do so?

5.6 ANSWERS
Self Assessment Questions
1. Any two areas may be given (from section 5.2)

2. The elements are similar chemically but differ in their ability to penetrate
cell membranes presumably due to their different ionic size. They also
differ in their transport mechanism and efficiency to activate enzymes.
Sodium ions are more abundant outside the cells, (blood plasma and in
the interstitial fluid surrounding the cells). They participate in transmitting
nerve signals and regulate the flow of water across cell membranes. The
transport of sugars and amino acids into cells is also influenced by Na+
ions.

But potassium ions occur at a higher concentration inside cells. They

also activate many enzymes and participate in the oxidation of glucose
to produce ATP. With sodium, K+ ions are also involved in the
transmission of nerve signals.

Terminal Questions
1. Bioinorganic chemistry involves the study and function of inorganic
substances in living systems. It also deals with the transport, speciation
and mineralization of inorganic materials and their use in mechanical
therapy and diagnosis.

2. Manganese and cobalt are present in enzymes. The enzyme arginase

have these metals and it has role in protein degradation. Co is present in
Vitamin B12. 87
Block 2 Bio-Inorganic Chemistry

3. Calcium ions play an important role in clotting of blood. Blood contains

prothrombin, a soluble protein. This is converted to the enzyme thrombin
by the action of prothrombin activator in the presence of Ca2+ ion.
Thrombin, again aided by Ca2+, now clots the blood by converting its
soluble fibrinogen into insoluble fibrin.

Prothrombin activator
Prothrombin Thrombin
+ Ca2+
+ Ca2+

Fibrinogen Fibrin

88
Unit 6 Functions of s-block Metals

UNIT 6

FUNCTIONS OF s-BLOCK
METALS

Structure
6.1 Introduction 6.3 Role of Mg2+ Ions in
Chlorophyll
Expected Learning Outcomes
6.4 Summary
6.2 Transport of Metals
+ + 6.5 Terminal Questions
Na -K Pump
6.6 Answers

6.1 INTRODUCTION
In the previous unit you were introduced to metals in biological system. Here
you will be studying about the functions of s-block metals pertaining to
bioinorganic chemistry especially about the Na+-K+ pump. After that the role
of magnesium ions in chlorophyll will be discussed. In the next unit you will be
learning about the role of iron in oxygen transport.

Expected Learning Outcomes

After studying this unit you should be able to:

 describe the Na+-K+ pump; and

 understand the role of magnesium ions in chlorophyll.

6.2 TRANSPORT OF METALS

Na+-K+ Pump
The sodium-potassium pump (also referred to as Na+-K+-ATPase pump). It is
found in the cell membrane of all animal cells and deals with transport of these
ions. The structure of the membrane and the presence of enzyme systems
that operate the ion pump determine the permeability properties of cell
membranes. The Na+-K+ pump coresponds to active transport i.e the transport
of ions through membranes against the natural tendency of ions to diffuse 89
Block 2 Bio-Inorganic Chemistry

towards lower concentration. Whenever there is a charge distribution

established across the membrane by active transport, an electrode potential
develops. Also, there is a difference in chemical composition of the fluids
inside and outside the cell.
Most animal cells have a higher concentration of potassium ions (K+) inside
the cell membrane than outside and a higher concentration of sodium ions
(Na+) outside the cell than inside. You will find that this concentration gradient
is not maintained in dead cells. Also if cells are cooled to 0 oC then the flow of
ions equalizes the gradient. The immediate source of energy to sustain this
disequilibrium is ATP, huge amounts of which are consumed to maintain this
gradient. Ion pumping occurs at a limited number of positions in the
membrane. Three Na+ ions are pumped outwards and two K+ inwards for each
molecule of ATP hydrolysed to ADP and inorganic phosphate. So, the ion in
the membrane, which is to be transported, moves to the other side and gets
released with the help of a carrier. The first clue came in 1957 when Jens
Skou disovered an enzyme of molar mass 100 kg mol-1 that hydrolyses
Mg-ATP only in the presence of Na+ and K+. It is to be noted here that ATP is
a simplified notation. In cells it is present as Mg2+-complex, the structure of
which is given in Fig. 6.1 below.
NH2
N
N
O O
N
N  P
O O O
HO O
O O 2+
P P Mg
O
HO
O O
2+
Fig. 6.1 : Structural formula of Mg -complex of ATP.

The ability of the enzyme correlates quantitatively with the extent of ion
transport. Another observation came that this enzyme ATPase is
phosphorylated only in presence of Na+ and Mg2+ ions. Moreover the
phosphorylated product is hydrolysed if K+ are present. These observations
are summarized by the cycle of enzyme reactions shown in Fig. 6.2, it has
been shown that the enzyme undergoes a conformational change during
phosphorylation. E1 and E2 are two enzyme conformation. Using these clues a
mechanism for the sodium-potassium pump was proposed as shown in
Fig. 6.3.
3Na+(cyto)
2K+(cyto)
E1
ATP

E1(3Na+)
E2(2K+)
Mg-ATP
P
Mg-ADP
H2O
+
E1P(3Na )
E2P(2K+)

E2P
2K+(ext) 3Na+(ext)

90 Fig. 6.2 : The cycle of enzyme reactions that accomplish sodium pumping.
Unit 6 Functions of s-block Metals

+ +
Fig. 6.3 : The Na -K pump (The sodium-potassium pump moves toward an
+ +
equilibrium state with the relative concentrations of Na and K shown
at left).

ATP and three Na+ bind to the inside of the membrane and the enzyme (E1)
gets phosphorylated. The product undergoes a conformation change –
eversion (which is like motion through a revolving door) that brings the bound
Na+ to the outside of the cell membrane. There the three Na+ are replaced by
two K+ ions. The attachment of K+ triggers dephosphorylation and loss of ATP
induces a conformational change that carries the K+ to the interior of the cell
where they are released. This process builds up a charge gradient because
three Na+ are expelled for two K+ incorporated and so there is a net loss of
positive ions, thus the resting membrane potential of the cell becomes slightly
more negative. The overall reaction can be summarized by equation 6.1.

3Na (in) 2K  (out) Mg2 (in) E - ATP  H2O 

… (6.1)
3Na (out) 2K  (in) Mg2 - ADP HPO24  H  E

Kinetic studies have confirmed that Mg-ATP is the substrate and free ATP
binds to the enzyme and inhibits its activity. Cleavage of the phosphoryl group
from ATP proceeds via, an SN2 mechanism forming a 5 coordinate unstable 91
Block 2 Bio-Inorganic Chemistry

intermediate which breaks down completing hydrolysis. An observation

supporting this is that minute (10-9 mol/L) constants of vanadate (VO34 )
inhibits the operation of ATPase. VO34  binds to ATP and the five coordinate
species formed does not break down, conformational changes do not occur
and K+ remain on the outside of the cell membrane.

The novel feature of active transport from an inorganic stand point is the
selectivity of the complexation between K+ and Na+. You will not find this in
simple systems. The question here is that why the enzyme is selective first to
Na+ and then to K+ in different conformations. Group I cations are not strongly
complexing and selectivity in simple system is based upon coulombic forces,
differences in ionic radii and hydration enthalpy. Crown ethers are
biochemically active ligands, increase permeability of cell membrane towards
cation and show specificity (for example 18-Crown 6 (18C6) favours K+ over
Na+ by factors of nearly 100). This reason of selectivity may be the reason that
a ligand can adopt conformation providing chelating site of right size. Bases
stronger than water will bind preferably to the smaller, harder acid Na+ and
vice versa. Displacement of water from the softer K+ is easier. However such
complexes where bases are weaker than water will be very weakly bound.

Two kinds of border guards control the movement of ions across cell
membranes, namely ion channels and ion pumps. The ion channels allowing
the passage of ion through membranes are typically constructed from protein
polymers in helical conformation. These helices form the walls of the channels
The selective pm and their precise conformation produce specific geometrical arrangement of a
+
passage of large K number of potential ligand-metal binding sites.
ions (ionic radius of
130 pm) over smaller Channels for Na+ transport are permeable to H+ whereas K+ channels are not.
+
Na ions (100 pm) is a These facts can be explained by assuming that Na+ channels are hydrated
special feature of the and larger. The more strongly hydrated Na+ moves into the sites inside the
potassium channel. channels while exchanging water ligands to bind to the water molecules of the
channel in turn. Na+ channels accept ions by coordination to H2O molecules
inside the channel. These water molecules can transport H+. The K+ channels
are formed from ligand sites of protein and are dehydrated.

Thus you have learnt in this section that the main function of the sodium-
potassium pump is to transport sodium ions out of the cell and potassium ions
into the cell. As a result, the most important purpose served by this is that the
neurons can function in this sort of condition. Also the cell’s membrane
potential becomes stable due to the differences in ion concentration.

In this context you should also understand the terms active transport and
passive transport. Active transport gets powered by ATP and results in
movement of ions inside the cell against the electrochemical gradient, with the
help of a special membrane bound protein. After this establishment of an
electrochemical gradient inside the cell, a cotransporter uses the same energy
to bring in another ion against the concentration gradient. Passive transport
involves specific channels for particular ions and carriers may bind to specific
proteins and undergo conformational changes to deposit the bound protein
inside the cell, which is known as facilitated diffusion. So this sort of passive
diffusions takes place along the electrochemical gradient without the use of
92 energy where ions move across the membrane through protein channels.
Unit 6 Functions of s-block Metals

Before moving to the next section, please try to solve the following SAQ.

SAQ 1
Give the general principle of the sodium-potassium pump.

6.3 ROLE OF Mg2+ IONS IN CHLOROPHYLL

In this section we are going to study the role of Mg2+ ions in chlorophyll. For
Porophine is a ring or
this, we are essentially going to study about photosynthesis. Photosynthesis four pyrrole rings
involves a series of complex reactions in the chloroplasts of plants with the linked by methine
overall conversion of water and carbon dioxide to carbohydrates and groups.
dioxygen. The overall reaction may be written as

xCO2 + xH2O (CH2O)x + xH2O … (6.2) Porphyrin is any class

of heterocyclic
All the oxygen comes from oxidation of H2O. as shown by the use of labelled compounds
containing four
pyrrole rings in a
square. It is present
*
xCO2 + 2 xH2O * + xH O
(CH2O)x + xO … (6.3)
2 2 in a form with a metal
atom in its cavity at
The reaction is endothermic to the extent of about 469 kJ per mole of CO2 and the centre, for
is initiated by visible solar radiation in the range 600-700 nm. A part of the example, in
series of reactions may proceed in dark as well. hemoglobin with iron.

Plants absorb light with the help of chlorophylls which are magnesium
porphyrin complexes (Fig. 6.4). Other variants of chlorophylls are also known
in different algae.

Fig.6.4: The chemical structure of (a) Chlorophyll a, (b) chlorophyll b,

Chlorophyll a contains methyl group (-CH3 ) and chlorophyll b contains
aldehydic group (-CHO) instead.

The molecules are capable of absorbing photons around 700 nm and transmit
the energy on to other species in the reaction sequence. The ability to absorb 93
Block 2 Bio-Inorganic Chemistry

photon may be due to the conjugated polyene structure of the porphyrin ring
system (Fig. 6.4). Some chlorophylls (actually chlorophyll-protein complexes)
act as antenna for solar radiation – also called light harvesting centres (LHC),
which pass the photon energy in the reaction site through excitation transfer
within picoseconds. Other pigments like carotenoids may also be present in
LHC; which respond to radiations not absorbed by chlorophylls.

The overall photosynthetic conversion is the outcome of a series of complex

reactions and involves light absorption at two different regions of wavelength –
photosystem-I (PS I) and photosystem-II (PS II). PS-I is based on a special
form of chlorophyll-a having absorption maxima at 700 nm (P-700) while PS II
utilizes another form of chlorophyll-a having maxima at 680 nm (P-680).

Photosystem-II: A manganese cluster catalyzes the oxidation of water to

oxygen; the metalloenzyme is called oxygen-evolving complex (OEC):

+
2H 2O O2 + 4H + 4e … (6.4)

The electrons are excited photochemically by P-68O to the primary electron

acceptor pheophytin. It is then transferred to photosystem I via the
intermediacy of (i) cytochrome b3, (ii) plastoquinone (iii) Rieske’s Fe-S protein
(iv) cytochrome f and (v) plastocyanin.

Photosystem-l: The electron received from PS-II is further excited by P-7OO

and is ultimately used up in the reduction of CO2 via (i) an Fe-S center (ii)
Ferredoxin NADP reductase and (iii) NADPH. The electron flow also results in
the formation of ATP from ADP.

Fig.6.5 : Schematic representation of the main steps in photosynthesis

(Z-scheme).

The main reactions which have been schematically represented as in Fig. 6.5
94 may be summarily represented as
Unit 6 Functions of s-block Metals

PS-II : 2H O +
2 O 2 + 4H + 4e ] x 6

PS-I : 2NADP + + 2H ++ 4 e 2NADPH ] x 6

Dark reaction : 12NADPH + 12H + + 6CO +

2 12NADP + 6H 2O + (CH2O)6
Net reaction : 6H 2O + 6CO 2 6O 2 + C 6H 12O 6
… (6.5)

The Mg2+ ion serves at least three purposes:

(i) it maintains the rigidity of the macrocyclic structure of chlorophyll and

minimizes molecular vibrations; hence energy is not readily lost by
thermal vibrations.

(ii) The H2O molecules coordinated to it mediates H-bonding between

adjacent chlorophyll molecules in a stack.

Before moving to the next section, please try to solve the following SAQ.

SAQ 2
What purpose does the magnesium ions serve in the process of
photosynthesis?

6.4 SUMMARY
In this unit you have studied about the functions of s-block metals pertaining to
bioinorganic chemistry especially about the Na+-K+ pump. Also, the role of
magnesium ions in chlorophyll were discussed.

6.5 TERMINAL QUESTIONS

1. Explain the Na+-K+ pump with a suitable diagram.

2. Give the main reactions in photosynthesis.

3. In which proteins in an adult human iron storage and and transport takes
place?

6.6 ANSWERS
Self Assessment Questions
1. The general principle of the sodium-potassium pump: Ion pumping occurs
at a limited number of positions in the membrane. Three Na+ ions are
pumped outwards and two K+ inwards for each molecule of ATP
hydrolysed to ADP and inorganic phosphate. So, the ion in the
membrane, which is to be transported, moves to the other side and gets
released with the help of a carrier. These observations are summarized by
the cycle of enzyme reactions (provide Fig. 6.3 here), it has been shown
that the enzyme undergoes a conformational change during
phosphorylation. E1 and E2 are two enzyme conformation. Using these
clues a mechanism for the sodium-potassium pump was proposed. 95
Block 2 Bio-Inorganic Chemistry

2. The Mg2+ ion serves at least three purposes:

i) it maintains the rigidity of the macrocyclic structure of chlorophyll

and minimizes molecular vibrations; hence energy is not readily lost
by thermal vibrations.

ii) The H2O molecules coordinated to it mediates H-bonding between

adjacent chlorophyll molecules in a stack.

Terminal Questions
1. The permeability properties of cell membranes depend upon the structure
of the membrane and the presence of enzyme system that operate the ion
pump. The Na+-K+ pump coresponds to active transport i.e the transport of
ions through membranes against the natural tendency to diffuse towards
lower concentration. The charge distribution established across the
membrane by active transport result in an electrode potential as well as
difference in chemical composition of the fluids inside and outside the cell.
Most animal cells have a higher concentration of potassium ions (K+)
inside the cell membrane than outside and a higher concentration of
sodium ions (Na+) outside the cell than inside. Three Na+ ions are
pumped outwards and two K+ inwards for each molecule of ATP
hydrolysed to ADP and inorganic phosphate. The simplest concept is that
of a carrier in the membrane which can combine with the ion to be
transported, carry it to the other side and release it. (Explain with the help
of Fig. 6.3)

2. The main reactions may be summarily represented as

PS-II : 2H O +
2 O2 + 4H + 4e ] x 6

PS-I : 2NADP + + 2H ++ 4 e 2NADPH ] x 6

Dark : 12NADPH + 12H + + 6CO +

2 12NADP + 6H 2O + (CH2O)6
reaction
Net : 6H 2O + 6CO 2 6O 2 + C 6H 12O 6
reaction
3. For Fe-transport- Transferritin, for Fe-storage- Ferritin.

Source of Figures
Fig. 6.4
[Link]
giers)/Unit_3%3A_Plant_Physiology_and_Regulation/13%3A_Photosynthe
sis/13.05%3A_The_Light-dependent_Reactions,CC-BY-SA license)

Fig. 6.5
[Link]

96
Unit 7 Role of Iron in Oxygen Transport

UNIT 7

ROLE OF IRON IN OXYGEN

TRANSPORT

Structure
7.1 Introduction 7.4 Summary

Expected Learning Outcomes 7.5 Terminal Questions

7.2 Hemoglobin 7.6 Answers

7.3 Myoglobin

7.1 INTRODUCTION
In the previous unit, you have learnt about the functions of s-block metals
pertaining to the living system especially about the sodium-potassium pump.
The role of magnesium ions in chlorophyll has also been discussed. In this last
unit of the block on bioinorganic chemistry you will be learning about the role
of iron in oxygen transport. Hemoglobin and myoglobin will be discussed in
detail as they are the best known iron porphyrin compounds. They are
metalloproteins and are special as they serve the prupose of oxygen carriers.
In this way, units 5, 6 and 7 will give you an insight of bioinorganic chemistry.

Expected Learning Outcomes

After studying this unit you should be able to:

 describe the salient features of hemoglobin; and

 discuss the special characteristics of myoglobin alongwith its structure.

7.2 HEMOGLOBIN
The commonest examples for bioinorganic chemistry in the context of carrying
oxygen to the cells is hemoglobin. Before we learn about hemoglobin we
should study about iron and its applications in the biosystem. 97
Block 2 Bioinorganic Chemistry

Even though iron is essential in small amounts for both plants and animals,
large amounts of it is toxic. The human body contains nearly 4 g of iron (Fe)
and it is the most important trace element, being involved in several processes
as below:

(i) As an oxygen carrier in blood of birds, mammal, fish (hemoglobin)

(ii) For oxygen storage in muscles and tissue (myoglobin)
(iii) As an electron carrier in plants, animals, bacteria (cytochromes) and
for electron transfer in plants and bacteria (ferredoxin)
(iv) Storage and scavenging of iron (Fe) in animals (ferritin and transferrin)
(v) As nitrogenase (enzyme in nitrogen (N2) fixing bacteria)
(vi) As a number of other enzymes like aldehyde oxidase, peroxidase
(decomposing of H2O2), succinic dehydrogenase (aerobic oxidation of
carbohydrates).
An adult human contains about 5 litre of blood, each millilitre of which contains
5000 million blood cells; each cell contains 2.5  105 (0.25 million) hemoglobin
molecules. The normal concentration of hemoglobin in blood of males is 14-16
g/dL and in females it is 13-15 g/dL. As the red blood cells is of short life span
that is in the range of 100-120 days, about 1% of these hemoglobin molecules
is replaced daily.

As you see that hemoglobin is essential for oxygen transport, whereas

myoglobin is engaged in storage of oxygen in muscle tissues. It is used for
the controlled release for oxidative phosphorylation as and when necessary.
What is the commonality between hemoglobin and myoglobin? Well, the site
of reversible binding of oxygen in both is heme (or haem), which is an Fe-
protoporphyrin IX where iron is in the Fe2+ state.

In hemoglobin, a high-spin Fe2+ is coordinated by the four N-atoms of

porphyrin and the fifth coordination position is bound to nitrogen atom of an
imidazole group in the histidine residue.

Out of nearly 4 g of iron in the human body, 70% is in the form of hemoglobin,
the red pigment in the erythrocytes that make up nearly 95% of the dry volume
of red blood cells. Here you may note that most of the remaining iron is stored
as ferritin. The function of hemoglobin is to pick up oxygen at the lungs. The
arteries carry blood to different parts of the body e.g. the muscles, where
oxygen is required. Here oxygen is transferred to a myoglyobin molecule and
stored until it is required to release energy from glucose. When oxygen is
removed from hemoglobin molecules, it gets replaced by a water molecule.
Oxygenation of hemoglobin causes release of H+ from a side chain.

Indirectly this helps to remove carbon dioxide ion from the tissues, since it is
converted to the more soluble bicarbonate ion, that is HCO 3 .

CO2  H2O  HCO3-  H .

+
As deoxygenated hemoglobin absorbs the H , the equilibrium shifts towards
98 the right and HCO 3 is transported between folds of the globin chain and there
Unit 7 Role of Iron in Oxygen Transport

is no definite binding between hemoglobin and hydrogen carbonate. Thus, it

acts as a carbon dioxide carrier by solubllising carbon dioxide as hydrogen Hemoglobin binds
carbon monoxide
carbonate. The transportation of carbon dioxide is by the amino acid side
more strongly than
chain and not by heme group and the blood returns to the heart via the veins. oxygen.
It is pumped to the lungs where hydrogen carbonate is converted in carbon
dioxide and the air is exhaled. The blood takes up oxygen and the process is
repeated.

The most striking feature regarding hemoglobin acting as an oxygen carrier is

the complete reversibility of the process. If the oxygenated complex was too
stable then huge amount of energy would be released in the lungs. Then
there would be less energy left by the time oxygen is released in the muscles.
The oxygenated form is called oxyhemoglobin and the deoxygenated form
deoxyhemoglobin. The transfer of oxygen is remarkable as it involves Fe2+ and
not Fe3+. Ligands like CO, PF3, CN- bind with hemoglobin irreversibly. These
ligands therefore prevent oxygen transfer and may result in death.
CH=CH2

CH=CH2

Fig. 7.3: Schematic diagram of haemoglobin showing (a) the heme, α chain 1, α
Protoporphyrin IX
chain 2, β chain 1 and β chain 2. (b) Structure of heme (haeme)
containing a protoporphyrin IX, with iron at its centre. consists of four
pyrrole rings to which
Molecular mass of hemoglobin has a molar mass of nearly 65,000. It is a four methyl, two
conjugated protein made up of four subunits which are four heme units. Each propionyl and two
vinyl groups are
subunit comprises a porphyrin complex (prosthetic group) heme which
attached.
consists of Fe2+ bound to four nitrogen atoms and it is associated with a
globular protein called globin. Heme belongs to the class of metal porphyrins
or metalloporphyrins. All hemoglobins carry the same heme as the prosthetic
group which is iron protoporphyrin IX as shown in Fig. 7.3a. The basic frame
work of all porphyrin is built of four pyrrole rings linked together in a larger ring
like structure called porphin. By substitution of the eight hydrogen atoms about
the edge of the porphin structure with various side chain and groups gives
various porphyrin found in nature. Heme contain –CH3 in 1,3,5 and 8,
propionyl (CH2CH2 COOH) in 6, 7 and vinyl in 2,4 positions. Each
hemoglobin molecule has four heme groups bound to globin on the surface as
shown in Fig. 7.4. For each heme unit of hemoglobin the four square planar
positions of Fe2+ are occupied by the four nitrogens of the porphyrin ring with
the fifth coordination site of Fe2+, away from the ring occupied by the
imidazole nitrogen of a histidine residue (proximal histidine) of the protein 99
Block 2 Bioinorganic Chemistry

(globin). Globin is made up of polypeptide chain of 141 (alpha) and 146 (beta)
amino acids. The vacant sixth site, below the ring is occupied by a loosely
held water molecule which is replaced by oxygen on oxygenation.

Fig. 7.4: Heme linked to the protein chain (GLOBIN).

The heme is further linked to the protein chain by condensation through the
–COOH end of the two propionyl groups. Fe2+ in hemoglobin is in the high spin
state but in oxyhemoglobin it is low spin. This highly complex molecule is
entirely synthesised in the living system by series of condensation reactions
beginning with glycine and succinate. The tetrahedral arrangement of
hemoglobin subunits gives the tight spherical appearance. Each individual
polypeptide is folded in a way so that the exposed surface has the maximum
of polar residues and the non-polar interactions remain internal. Thus this
large protein becomes water soluble. Heme is held within the cleft of the
associated protein molecule namely globin with 150-160 amino acid residues.
The polypeptides in hemoglobin are held together by hydrogen bonding,
hydrophobic interactions and multiple ionic interactions. The α-chain consists
of 141 amino acid residues in linear sequence. The β, ϒ and δ chains consist
of 146 amino acids. In a helical arrangement, the 75% of the amino acids are
in α or β-chains. The haemoglobin tetramer has two identical dimers (α, β) 1
and (α β) 2 [dimer1 and dimer2] which are held together by polar bonds. There
is extensive hydrophobic interaction between the porphyrin part and the globin
molecule, besides the covalent interaction between Fe2+ and the proximal
histidine. This arrangement is considered as its quaternary structure of
hemoglobin as depicted in Fig. 7.3.

In the free state octahedral Fe2+ in heme has its two coordination positions
above and below the plane occupied by water molecules. The magnetic
moment of heme shows presence of four unpaired electrons characetrisitic of
a weak field octahedral complex of d6 Fe2+. It is very sensitive to oxygen and
readily combines with it to form a labile Fe2+ oxygen complex (oxy-heme)
intermediate which changes to the Fe3+ porphyrin complex which is hemin as
shown in Fig. 7.5.
OH 2 +
O2 OH 2

N II N
O2 N N N III N
Fe
Fe Fe
N N
N N N N
OH 2
OH 2 OH 2
Heme Oxy-heme Hemin
100 Fig. 7.5: Conversion of heme to hemin.
Unit 7 Role of Iron in Oxygen Transport

Now, let us understand how this conversion of heme to hemin is prevented in

hemoglobin. In hemoglobin, the Fe2+ heme is enclosed in a hydrophobic
pocket globin, which provides a medium of low dielectric constant and
prevents water molecules or any polar solvent to occupy the sixth position
which is occupied by a protein residue. It does not allow simultaneous
coordination of oxygen and water which seems to be necessary for electron
transfer to transform Fe2+ to Fe3+. The access to the iron is reduced as the
protein chain is folded about the heme groups. In the absence of globin, Fe2+
heme would get irreversibly oxidized to Fe3+ which would not be able to
transport oxygen. Oxidation is irreversible and a -peroxo and finally -oxo
Fe2+ heme dimer is formed as shown below:

Fe2+ heme  O2  Fe2 + heme O-2 (superoxide) 

Fe3 + hemeO -2  Fe2+ heme  Heme Fe3 + O  O Fe3 +heme


Heme Fe3 +  O  Fe3 + heme

This complex cannot carry oxygen reversibly and its formation would stop the
transport of oxygen to cells and tissues and all body processes would come to
a halt. The role of the globin chain is both chemical and steric.

 It provides a medium of low dielectric constant. Whereby charge

separation as in -peroxo and -oxo dimer is not possible.
 It acts as a carrier of heme and stabilises the heme–oxygen complex such
that oxidation of Fe2+ in heme is prevented.
 It does not allow simultaneous coordination of oxygen and water which is
a prerequisite for oxidation of Fe2+ to Fe3+.
 The heme is deep-buried in the protein fold and this prevents the
peripheral attachment to Fe2+ and thus prevents its oxidation.

 The role of the fifth ligand i.e. histidine is to provide optimum electron
density on iron. If the fifth ligand is highly basic then the electron density
on Fe2+ increases and when oxygen is taken up the ineraction is so strong
that the reaction is irreversible and hemoglobin no longer can act as an
oxygen carrier. If the fifth ligand is weakly basic then the electron density
on Fe2+ is not increased sufficiently and the bond between iron and
oxygen is not strong and dissociation is rapid. Hence the fifth ligand
should have sufficient basicity to maintain optimum bond strength
between iron (Fe2+) and oxygen.

There is another histidine situated in the oxygen binding pocket on the side
away from the coordinated imidazole base. This is called the distal histidine.
Neutron diffraction studies indicate the formation of a hemoglobin bond
between the coordinated dioxygen molecule and the N-H proton of the
histidine residue: This FeOO…..HN unit is accommodated very well as
there is the angular bend at the coordinated dioxygen atom. This is
established by crystallography. The double bond angle is equal to 120o. This
coordination is favoured over straight as in CO due to the distal histidine. In
carbon monoxide adduct of hemoglobin, X-ray data reveal a distorted structure 101
Block 2 Bioinorganic Chemistry

in which the FeCO angle is either bent or tilted off to the proximal histidine
NFe (natural tendency for NC=O bond is linear as in carbonyls) axis to
avoid steric clash with the distal histidine bond. So you see, how intelligently
nature created the Fe-porphyin centre so that oxygen binds to it rather than
the toxic carbon monoxide. Here you must note that carbon monoxide often
binds more tightly to metal complexes than does oxygen.

Next let us understand the Perutz mechanism in the context of oxygenation of

hemoglobin. An interaction between an oxygen molecule and a heme group
can affect the position of the protein chain attached to it, which in turn affects
other protein chains through hydrogen bonds. The key or trigger to the Perutz
mechanism is the high spin Fe2+ atom in an oxygenless heme. The
hemoglobin has two conformational states. The deoxygenated form is more
tense and is called the Tense (T) form, whereas the oxygenated form is the
Relaxed (R) form (methemoglobin). The T state is controlled by the subunit-
subunit interactions, but R state is more flexible due to the ability of oxygen
binding. The radius of high spin Fe2+ is nearly 78 pm. The Fe-N bond distance
in heme is nearly 218 pm. The size of the central hole is nearly 200 pm and so
the Fe atom in the T form must lie above the plane of the heme group. The low
spin Fe2+ is almost 17 pm smaller than the high spin Fe2+; the Fe-N bond
distance is almost exactly 200 pm and low spin Fe2+ fits smugly into the hole.
Coordination with oxygen causes the Fe atom to drop into the plane of the
heme group and the Fe2+ changes from high spin in T form to low in R form.
Hemoglobin is made up of four subunits and when one subunit takes up an
oxygen molecule, the Fe2+ contracts and fits into the plane of the ring. As this
happens, the histidine residue bound in the fifth coordination site moves with it
as shown in Fig. 7.6.

Fig. 7.6: Structural change in hemoglobin on reversible binding of O2.

The distribution of electrons in the “Tense “ and “Relaxed” form, according to

the octahedral symmetry of crystal field theory will be as following:

Relaxed form
eg

t2g

102
Unit 7 Role of Iron in Oxygen Transport

Since high spin Fe2+ contains two electrons in antibonding orbitals, it is larger
than low spin Fe2+ which has all electrons in bonding orbitals. The Fe-N bond
length is related to bond order. In T form two electrons are present in eg
orbitals which decreases the bond order and increases the bond length.
Oxygen has  acid properties and is a strong ligand. It induces large crystal
field splitting when coordinated to Fe2+; thus the electrons are paired up in low
lying t2g orbitals resulting in a low spin complex.

Mechanical depiction of shrinkage of Fe2+ and movement into the porphyrin

ring causes conformational changes in the globin chain. Since this chain is H-
bonded to the other three units, it changes their conformation too and
enhances their ability to attract oxygen. This phenomenon is called
cooperative effect (allosteric effect). When oxygen pressure is high as in
lungs, the oxygen uptake by hemoglobin increases due to cooperative
interaction. In a similar way when the blood reaches the muscles where the
oxygen pressure is low, once one oxygen is released, the others are released
even more readily and the cooperative effect is in reverse order. The
deoxygenated form of hemoglobin is also said to have a domed structure as is
clear from the Fig. 7.7.

Fig. 7.7: The domed structure of the deoxygenated hemoglobin.

The affinity of hemoglobin for oxygen decreases with decreasing pH, i.e.
oxygenation is pH dependent and is favoured at high pH. This is referred to as
Bohr effect. There is long range electrostatic interaction of the type
 
COO …….. NH4 and H-bonded interaction of the type OH…… OOC
between the globin chains. Oxygenation of hemoglobin involves in breaking of
these interactions and is pH dependent. With increase in pH, breaking of
bonds gets favoured and oxygen uptake increases; while at low pH
electrostatic interactions are strengthened resulting in lowering of oxygen
uptake. So, in region of low pH, hemoglobin has a tendency to release
oxygen. In the lungs, the pH is 7.4. The pH at which oxygen binding affinity is
lowest is 6-6.5 and the working tissues provide this pH. Blood is well buffered
by the presence of carbon dioxide. Carbon dioxide dissolves in water to form
carbonic acid, assisted by the enzyme carbonic anhydrase.

CO2  H2O ⇌ H2CO3 ; H2CO3 ⇌ H  HCO3- …(7.1)

In the muscles there is a good concentration of carbon dioxide and thereafter

hemoglobin takes them up. The terminal amine group of protein picks up the
H+ produced (as a result of Eq. 7.1). When the supply of oxygen goes down,
glucose undergoes incomplete oxidation to lactic acid. Thus the pH is lowered 103
Block 2 Bioinorganic Chemistry

leading to further release of oxygen from oxyhemoglobin. So in this section,

you have learnt about the structure and functions of hemoglobin. Before
moving to the next section try to answer the following SAQ.

SAQ 1
Explain with the help of a suitable figure, how the binding of oxygen takes
place in oxyhemoglobin.

7.3 MYOGLOBIN
This is the last section in this block on bioinorganic chemistry where
myoglobin, the oxygen carrier for getting oxygen into the cell for glucose
oxidation will be taken up. It is the pigment in muscles resembling hemoglobin
in its function. It is the simplest example of a respiratory heme protein and its
molecule consists of only one polypeptide chain and one heme group. It has a
molar mass of nearly 17000 and acts as an oxygen reservoir within the muscle
fibre. It has a higher capability for oxygen binding than hemoglobin and can
get oxygenated at any given pressure.

The molecule of myoglobin has a capability to bind oxygen. This capability is

acquired by the presence of heme, a non-polypeptide prosthetic group
consisting of protoporphyrin IX and a central iron atom. This complete
structural arrangement is known as protoheme group as shown in Fig. 7.8 (b).
This protoheme group is stabilized by histidine residue above (His 64) and
below (His 93) as shown in Fig.7.8 (a). The two histidine residues located in
the interior part of the molecule also facilitate the binding of iron and oxygen.

(a) (b)
Fig. 7.8: (a) Schematic diagram of myoglobin, (b) Structure of myoglobin showing
the protoheme group.

Hemoglobin is a tetramer with electrostatic interaction between different units

resulting in a constraint in structure, thus the overall rate of binding with
oxygen is less in hemoglobin than with myoglobin and this explains why
myoglobin can take up oxygen at low pressure. The function of hemoglobin is
to pick up oxygen at high oxygen partial pressure in the lungs and carry it
without loss through the blood and release it to myoglobin. So, myoglobin
104 must be having greater affinity for oxygen than hemoglobin at low pressure.
Unit 7 Role of Iron in Oxygen Transport

The loading tension refers to the tension of saturation of 95% of pigment with
oxygen. The value (pO2 in tissues is equal to 40mm of Hg) for hemoglobin is

Fig. 7.9: Saturation of hemoglobin with oxygen against partial pressure of oxygen.

73 mm hemoglobin. In tissues pO2 is low, so hemoglobin can give up oxygen.

The oxygen saturation of hemoglobin as a function of pO2 is shown in Fig. 7.9.

The oxygen saturation curves for hemoglobin and myoglobin have different
shapes. It is hyperbolic for myoglobin and sigmoidal for hemoglobin as shown
in Fig. 7.10. The isotherm for myoglobin is expected of this shape as the
equilibrium is simple with one oxygen which binds to one iron. However the
isotherm for hemoglobin has a different shape as the binding of oxygen at one
subunit influences the oxygen binding to the other. The shapes of the oxygen
saturation curves for myoglobin and hemoglobin are explained in terms of this
equilibrium is shown in Fig. 7.10 where it shows the fractional oxygen saturation
 as a function of the oxygen partial pressure at pH 7.2.

Fig. 7.10: Oxygen saturation curves for myoglobin and hemoglobin.

[Mb 2 O 2 ]
Mb  O 2  MbO 2 ; K  …(7.2)
[Mb]p
p partial pressure O 2
 = fractional O2 saturation = ratio of concentration of myoglobin present as
oxymyoglobin (MbO2) to the total myoglobin concentration. 105
Block 2 Bioinorganic Chemistry

[MbO 2 ] K[Mb]p [Mb]Kp Kp

α    …(7.3)
[Mb]  [MbO 2 ] [Mb]  K[Mb]p [Mb](1  Kp) 1  Kp

 [Kp/(1  Kp)] Kp 1  K p  K p
Saturation factor =   
1  1  [Kp/(1  Kp)] 1  Kp 1  Kp

Kp 1  Kp
   Kp …(7.4)
1  Kp 1

This Kp shown in Eq. 7.4 is the saturation capacity, which when equal to 1
means that there is 100% saturation. In case of hemoglobin, the curve can be
reproduced if dependence of  on partial pressure is represented as pn.

[Hb(O2 )n ]
Hb  nO 2 Hb(O2 )n K …(7.5)
[Hb]pn

[Hb(O 2 )n ] K[Hb]p n Kp n
   …(7.6)
[Hb]  [Hb(O 2 )n ] [Hb]  K[Hb]p n 1  Kp n

α
  Kp n …(7.7)
1 α

If we take the logarithm of Eq. 7.7, we get

α
log  logK  n log p …(7.8)
1 α

α
The Eq. 7.8 is referred as Hill equilibrium. If log is plotted against log p
1 α
then the slope will be 1 for myoglobin as n = 1 and for hemoglobin it is
2.8 (n = Hill’s constant). This plot is called Hill’s curve. Hill’s constant refers to
the degree of cooperativity between different heme units. Since myoglobin is a
monomer it does not have the feature of cooperativity. Hemoglobin picks up of
oxygen in a stepwise manner. In this process, steric hindrance comes into
play and so the maximum number of oxygen picked up is reduced to three.

Difference between hemoglobin (Hb) and myoglobin (Mb)

 In terms of its structure, hemoglobin is made up of four subunits,

myoglobin has only one subunit

 Hemoglobin has constraint in structure due to electrostatic and other

interactions between different subunits, myoglobin has no such constraint.

 Hemoglobin picks up oxygen only at high partial pressure whereas

myoglobin can pick up oxygen at any pressure. The nature of oxygen
binding curve of hemoglobin is sigmoidal whereas in myoglobin it is
hyperbolic curve.

 The binding of oxygen to hemoglobin enhances binding of additional

oxygen to the hemoglobin molecule. In other words oxygen binds
cooperatively to hemoglobin whereas no such effect is noted in
106 myoglobin.
Unit 7 Role of Iron in Oxygen Transport

 The affinity of hemoglobin for oxygen is dependent on pH whereas that of

myoglobin is not affected by pH (Bohr Effect) .

 Dissociation of oxyhemoglobin is influenced by concentration of carbon

dioxide [CO2] and temperature (T). In muscular tissue both pressure of
CO2 and temperature are high. High pressure of CO2 results in low pH
and thus hemoglobin gives up oxygen readily.

 Affinity of hemoglobin for oxygen is controlled by 2,3-diphosphoglycerate

present in red blood cell (RBC). Total hemoglobin has less affinity for
2,3-DPG in comparison to adult hemoglobin and hence higher affinity for
oxygen. This facilitates transfer of oxygen from mother’s oxyhemoglobin
to fetal hemoglobin. Myoglobin is found in muscles.

So in this section, we have discussed about the structure and functions of

myoglobin and also compared them with those of hemoglobin. Now, answer
the following SAQ.

SAQ 2
Mention any three differences between hemoglobin and myoglobin.

7.4 SUMMARY
In this unit, you have studied the role of iron in oxygen transport. You have
learnt about the different features of hemoglobin and myoglobin and compared
them too. In this way, you have learnt all about bioinorganic chemistry with the
help of units 5, 6 and 7 of this block. This brings us to the end of this section of
inorganic chemistry.

7.5 TERMINAL QUESTION

1. What is the “cooperative effect” regarding the binding of oxygen with
hemoglobin?

2. Give the relevant equations for the equilibrium constant regarding the
oxygen binding by myoglobin.

7.6 ANSWERS
Self Assessment Questions

1. In oxyhemoglobin the plane of the porphyrin ring becomes nearer to

the iron which was out of plane. This serves as a trigger for the
cooperative binding of oxygen to the four subunits in hemoglobin.
When there is no oxygen, these subunits interact between themselves
so that the proximal histidine can not move the five coordinate iron
towards the plane of the porphyrin.
107
Block 2 Bioinorganic Chemistry

2. The difference between hemoglobin and myoglobin are:

[Link]. Hemoglobin Myoglobin

i) Hemoglobin takes up oxygen Myoglobin can take up

only at high partial pressure oxygen at any pressure

ii) Hemoglobin can take up Myoglobin can take up

oxygen in a particular pH oxygen at any pH

iii) Hemoglobin has four heme Myoglobin has only one

groups per molecule heme group per molecule

Terminal Questions
1. When dissolved oxygen is present hemoglobin undergoes certain
changes in its conformation. As one iron binds to an oxygen molecule,
the additional oxygen molecule can be much easily binded to the
hemoglobin because of certain changes in its molecular shape. So,
total four irons of the hemoglobin can take up one oxygen each, and
the equilibrium constant increase for each consecutive step. This
triggering of binding of oxygen is known as cooperative effect.
Similarly, the removal of one oxygen also helps in the removal of the
remainder oxygens.

2.  = fractional O2 saturation = ratio of concentration of myoglobin

present as myoglobin O2 to the total myoglobin concentration.

[Mb 2O 2 ] K[Mb]p [Mb]Kp Kp

α   
[Mb]  [Mb 2 ] [Mb]  K[Mb]p [Mb]  (1  Kp) 1  Kp

Kp 1  Kp
   Kp  saturation capacity   100%
1  Kp 1

Further Reading

1. Inorganic Chemistry (4th Edition) by James E Huheey, Ellen A Keiter,

Richard L Keiter & Okhil K Medhi, PEARSON.

2. Inorganic Chemistry (4th ed.), J.E Huheey, Keiter, Keiter and Medhi,
Pearson Education, 2006.

3. Chemistry of the Elements. N.N. Greenwood and A. Earnshaw,

Pergamon.

4. Inorganic chemistry by D. F. Shriver, P. W. Atkins and C. H. Langford,

Oxford University Press.

5. An Introduction to Inorganic Chemistry by Purcell and Kotz.

6. Ramaprasad Sarkar, General and Inorganic Chemistry, NCBA.

7. R. L. Dutta, Inorganic Chemistry - Part-II - Chemical Elements & Their

Compounds, The New Book Stall.
108