METALLOPROTEINS
Metalloproteins are a class of biologically important macromolecules and account for nearly
half of all proteins in biology. They are responsible for performing some of the most difficult
yet important functions, including photosynthesis, respiration, water oxidation, oxygen
transport, electron transfer, oxygenation and nitrogen fixation. These diverse functions of
metalloproteins have been supposed to depend on the ligands from the amino acid,
coordination structures, and protein structures in the immediate vicinity of metal ions.
Huge amount of efforts have been devoted toward understanding the structure and
function of metalloproteins.
ELECTRON TRANSFER PROCESS
Metalloprotein which has a single type of redox cofactor can be classified into two general
groups: electron carriers and proteins involved in the transport or activation of small
molecules. It is assumed that proteins that function as electron transferases typically place
their prosthetic groups in a hydrophobic environment where in addition to ligand,
hydrogen bonds may be provided which help to assist in stabilization of both the oxidized
and the reduced forms of the cofactor. In order to minimize inner-sphere reorganization,
metal-ligand bonds remain intact upon electron transfer.
There are four classes of electron transferases
blue copper proteins, cytochromes, flavodoxins, and iron-sulfur proteins.
each of which contains many members that exhibit imp function and structural differences.
IRON SULPHUR PROTIEN
Iron sulphour proteins are proteins characterize by the presence of iron sulphour cluster
containing sulphour linked Di, Tri, and tetra iron centres in variable oxidation state. Iron
sulphur clusters are found in a variety of metalloproteins such as ferredoxins, as well as
NADH degydrogenase, hydrogenases & NADH dehydrogenase, hydrogenases &
nitrogenases etc. Iron sulphour clusters are best known for their role in the oxidation
reduction of electron transport in mitrochondria and chloroplasts.
Two iron sulphour protein are  Rubredoxins & ferredoxin
Rubredoxins:- The simplest of all iron-sulfur
proteins are the rubredoxins, which are primarily
found in anaerobic bacteria. Rubredoxins are small
proteins (6 kDa) and contain iron ligated to four Cys
sulfurs in a distorted tetrahedral arrangement. The
reduction potentials of iron-sulfur proteins are
typically quite negative, indicating a stabilization of
the oxidized form of the redox couple as a result of
negatively charged sulfur ligands. Ferredoxins [2Fe-
2S] are the other class of iron-sulfur proteins (10-20
kDa) which are found in plant chloroplasts and
mammalian tissue.
Ferredoxins: Basically, they re three of types 2Fe-2S , 4Fe-4S, 3Fe-4S
1).. 2Fe-2S protein (Fe2S2 Ferredoxin) = Acidic in
nature, each Fe atom is bonded to 2 S-arom of cys & 2
bridging sulphour ions.
Distorted tetrahedral structure.
2).. 4Fe-4S ferrodoxin – It has
cubane type structure , Four iron
ions & four sulphide ions placed at
the verticies of a cubane type
cluster.
3).. 3Fe4S Ferredoxin :- 3Fe centre ferrodoxin was
first recognized in nitrogen fixing bacteria, 3Fe ions
present in +3 oxidation state, Reduced from 2Fe+3 Fe+2
The optical spectra of all iron-sulfur proteins are very
broad due to numerous overlapping charge-
transfer transitions that impart red-brown-black colors
to these proteins.
In contrast, the EPR spectra of iron-sulfur clusters are quite distinctive, and have
significant value in the redox chemistry studies of these proteins.
CYTOCHROMES:- cytochromes are
proteins containing one or more
heme cofactors. These proteins are
generally classified on the basis of
heme type. There are three most
commonly
encountered types of heme: They
found in both plants and animals,
contain heme like prothestic group.
1).. Cytochrome A  Heme group has a
formyl group replacing a methyl group.,
they are capable of binding O2 &
reducing it. It is responsible for severe
toxicity of CN-
2).. Cytochrome B  The heme group is
similar to hemoglobin and myoglobin but
protein is not bonded to heme, These
complexes are involved in electron
transport, the pumping of protons to create
a proton-motive force (PMF). This proton
gradient is used for the generation of ATP.
These complexes play a vital role in cells.
3).. Cytochrome C  Protein is attached to
heme through covalent linkage,
Cytochrome c is the most stable and
abundant member of the class. While most
heme proteins are attached to the prosthetic
group through iron ion ligation and tertiary
interactions, the heme group of cytochrome
c makes thioether bonds with two cysteine
side chains of the protein
FERRITIN
Ferritin, an iron-storage protein found in animals and plants, contains the largest cluster
currently known, an ordered aggregate {(FeOOH)8(FeO H2PO4)}n containing up to 4,500
Fe(III) atoms. The cluster occurs within a protein cavity roughly 80 angstroms in diameter.
Proteins in which heavy metal ions are bound directly to some of the side chains of
histidine, cysteine, or some other amino acid are called metalloproteins.
Red blood cells need iron to form normally and carry oxygen around your body. Other
parts of your body, such as your liver, bone marrow, and muscles, also need iron.
Low levels of ferritin lead to iron-deficiency anemia. This means you have too few red
blood cells. Iron deficiency can come from a poor diet or blood loss. Or your body may have
trouble absorbing iron from food.
High levels of iron can damage your joints, heart, liver, and pancreas. Too much iron is
most often caused by an inherited disease called hemochromatosis. Many people with this
disease never have any symptoms, especially women who lose iron through menstruation.
Iron poisoning happens when a large amount of iron is taken in all at once. This happens to
children who accidentally take too many iron supplements.
ferritin, which contains 20 to 22 percent iron, is the form in which iron is stored in animals;
it has been obtained in crystalline form from liver and spleen.
BIOLOGICAL NITROGEN FIXATION NITROGENESE
Biological nitrogen fixation (Figure 4) plays an essential part in providing nitrogen for other Biological N2 fixation mediated by enzyme complex nitrogenese. The atmospheric
forms of life on earth, since it contributes about 60% of the total N2 fixed in the nitrogen is chemically inert hence to fix N2 energy rich conduction are required high
biogeochemical nitrogen cycle. BNF is therefore called a key for sustenance of agriculture temp or electrically discharge can supply the required activation energy, but the
and reduction of soil fertility decline. natural process of BNF deals diazotropes fix N2 under mild condition at ambient temp & pressure with the help of
an economic means for the reduction of environmental problems and improvement of the a complex enzyme with multiple redox centre known as nitrogenese.
internal resources. Three different type of nitrogenase
In this process microorganisms transform atmospheric nitrogen to ammonia, assailable by Molybdenum, Vanadium & Iron Nitrogenase
associated plants. These microorganisms need 16 moles of adenosine triphosphate (ATP) NITROGENESE ENGYME
for the reduction of each mole of nitrogen and they obtain this energy by oxidation of The most acceptable structure of nitrogenase enzyme is given by Molybdenum Nitrogenase the
organic molecules. Free-living microorganisms which are non-photosynthetic in nature, dominant hypothesis. Nitrogenase enzyme has two component .
receive these molecules from other organisms, while photosynthetic microorganisms, such Component 1st  it is known as iron protein, it has two identical subunit which bind Fe4S4 by
iron Sulphur bond of cystein residue (Cyst-97 & Cyst-132).
as cyanobacteria, use sugars synthesized in the process of photosynthesis.
FeSO4 centre
N2 + 8H2+ 16ATP ------> 2 NH3 + 2H2+ 16ADP + 16 Pi Mw  57000 - 73000
Associative and symbiotic nitrogen-fixing microorganisms obtain these compounds from Fe – Protein
their host plants’ rhizospheres. ( Figure – 1)
Asymbiotic Bacteria or free living bacteria- fixes 30 % of N2
Symbiotic Bacteria- fixes 70 % of N2 & live in a symbiotic relationship with plants of the These sub unit produce by Nitrogen - H – gene, The iron protein is reduced by fleodoxine &
legume family (e.g., alfalfa , soybeans) ferrodoxine, The Fe4S4 centre under goes a single olectrane redox process the redox form is
EPR active & oxidize form is di magnetic., This Fe4S4 centre can exist in two spin state D = ½ ,
3/2 ., This protein bind two molecule of Mg ATP, a left b/w the two sub unit as ATP binding site.
(II) Fe-Mo Protein :- This protein has both iron and molybdenum, It consist four sub unit
2α+2β, 2 α sub unit originated from Nitrogen – D – gene, 2β subunit originated from nitrogen -
K – Gene. , These protein have two type of centre a = P – Cluster, b = Fe-Mo co factor (M-Centre).
Fe-Mo Co-factor
P – Centre
( Figure – 2)
2β Sub unit 2α Sub unit
(Nit – K – Gene) ( Nit-D-Gene)
a).. P – Cluster  These are 4 in number and located at αβ sub unit interface. These are Fe4S4
like centre in their oxidised form they have high spin and have 7/2 spin. Each P-cluster has 8
iron atom and 7 sulphide., In the reduced form each cluster take the form of 4 iron 3 sulphor
Partial cube linked by a center sulfide ion each cluster is linked to protein throw 8 cysteinate
residue.
Function of P cluster :- The storage of electron, e transfer Fe-Mo co-factors for substrate
reduction.
b).. Fe-Mo co factor :- These are two in number and located at α – Sub unit.
Function of Fe Mo co factor :- It is sight of reduction of the substratr. this is the site of N2
fixation. this is the site substrate binding.
Figure – 1 (FeSo4 centre )
ATP -----------1e- energy-----------------------  ADP + Pi
Two kinds of nitrogen-fixing bacteria are known: free-living or non-symbiotic bacteria, Figure - 2 N2 + 3H2  2NH3
including the cyano bacteria (or blue-green algae) Anabaena and Nostoc and genera such as
Azotobacter, Beijerinckia, and Clostridium; and mutualistic or symbiotic bacteria such as Mechanism of nitrogenase action
Rhizobium, associated with leguminous plants (e.g., various members of the pea family), and Nitrogenase catalyzed the reduction N2 to ammonia the Fe protein., provides electrons with
high reducing power to Fe-Mo protein which uses these electrons to N2 to NH3., this transfer of
certain Azospirillum species, associated with cereal grasses. electron from iron protein is coupled to ATP hydrolysis by Fe protein.
1)… Nitrogen Fixation by Free-Living Bacteria
There are many heterotrophic bacteria which reside in ground soil and are able for fixation of
significant levels of nitrogen without the direct interaction with other beings. Examples for this
type of nitrogen-fixing bacteria include species of Azotobacter, Bacillus, Clostridium,
and Klebsiella. These organisms search their own source of energy either by oxidation of organic
molecules released by other organisms or from decomposition. Some free-living organisms have
chemolithotrophic capabilities which help them to utilize inorganic compounds as a source of energy.
Their contribution to global nitrogen fixation rates is supposed to be less than generally observed due
to the lack of suitable carbon and energy sources for these microorganisms.
Associative Nitrogen Fixation
Species of Azospirillum form close associations with several members of the Poaceae (grasses),
that includes agriculturally economic and vital cereal crops, such as rice, barley, oats, wheat and
corn. These bacteria fix appreciable amounts of nitrogen within the rhizosphere of the host plants. The
degree of N fixation is limited by various elements, including soil temperature, the power of the host
plant to provide a rhizosphere environment low in oxygen pressure, the availability of host
photosynthates for the bacteria, the competitiveness of the bacteria, and the efficiency of nitrogen
2)… Symbiotic Nitrogen Fixation
Many microorganisms fix nitrogen symbiotically by partnering with a host plant. Sugars are .. 8 time cycle to fix one N2 Fe-Mo Co-Factor
produced by plants via photosynthesis that are utilized by the nitrogen-fixing microorganism for the N2 + 3H2 2NH3
energy it required for nitrogen fixing. In exchange for these carbon sources, the microbe N2 + 8e- + 8H+ + 16ATP + 6H2O  2NH3 + H2 + 16ADP + 16Pi
provides fixed nitrogen to the host plant for its growth. Water fern Azolla’s which symbiosis with a In most di azotrophs these eight high potentials e- comes from reduced Ferridoxime by
cyanobacterium Anabaena azollae is a type of example for this type of nitrogen fixation. Even though photosynthesis process the overall process involved in the following steps
the symbiotic partners described above play a vital role in the worldwide ecology of nitrogen fixation, 1 electron deduction of Fe4S4 four center of iron protein initiate the reaction.
till date relationships between legumes and Rhizobium and Bradyrhi zobium bacteria are considered two molecules of magnesium atp bind to iron protein.
to be the most important nitrogen-fixing symbiotic associations. this binding of magnesium atp induces conformational changes that permit the iron protein to
Legume Nodule Formation combine with Fe-Mo protein to form a complex.
The symbiotic nitrogen-fixing bacteria invade into the host plant’s root system, where they
multiply and stimulate the formation of root nodules, enlargements of plant cells and bacteria in
intimate association. The bacteria then start to fix the nitrogen required by the plant present in the
nodules. Access to the fixed nitrogen helps the plant to synthesize leaves fortified with nitrogen which
can be recycled throughout the plant. This results in increased photosynthetic capacity, which in turn
yields nitrogen-rich seed. It is quite amazing to note the outcomes of legumes which have not being
nodulated, especially when nitrogen poor soil is used for the plant growth. The plants produced are
usually low in nitrogen content, and yield very little seed.
[ Crop  Nodulating Bacteria ] Alfalfa  Sinorhizobium melilotii, Beans  Rhizobium
legumninosarum biovar phaseoli and Rhizobium tropici, Clover  Rhizobium leguminosarum biovar
trifolii, Lotus  Mesorhizobium loti, Peas  Rhizobium leguminosarum biovar viceae, Soybean 
Bradyrhizobium japonicum, Bradyrhizobium elkanii.
Na+/K+ pump
Sodium & Potassium are proteins in the form of Salts of inorganic acids in plants as well as animals.
found in the form of salts of organic acids, Sodium major extra cellular Cation and potassium is a
cellular cation. The biological contribution or biochemistry of sodium and potassioum as follows
1). . Hydration & Osmotic pressure control :- The main functions of sodium and potassium ions in The electron is transferred from iron protein to FE-MO protein that is coupled to atp hydrolysis
the body are various. the normal osmotic pressure of the fluids has to be maintained through which the binding and hydrolysis of triggers conformational changes that moves iron protein close to
a Fe-Mo protein so that it can effectively transfer its electron to the site of nitrogen reduction.
excessive loss of fluids is prevented.
2).. Acid-Base equilibrium control :- Sodium ions in extracellular fluids and potassium in this cycle repeated eight times following electron transfer the iron protein disassociate from
Fe-Mo protein this is the RDS.
intracellular pollutes they form salts with the weak acid present there these salts act as buffers and the
ph of the fluids in the body is maintained. In Fe-Mo protein the electron first reaches in P cluster then transfer to Fe-Mo Co-factor where
Mo is present in +6 oxidation state but after reduction it will be in +2 oxidation state.
3).. Convection of CO2 :- Na+ K+ and Cl- and ions play a major role in the convection of gaseous CO2.
4).. Blood viscocity control :-- Sodium and potassium ions maintain globulins in blood plasma and Fe-N≡N  Mo(-N=N-) 2e-/-2H+ Mo(-NH-NH-) Mo(iv)  NH=NH 2e-/2H+ NH2-NH2
2NH3  2e-/2H+
plasma moisture controls the amount of water secretes occurs.
THE SODIUM POTASSIUM PUMP :- The sodium-potassium pump (Na,K-ATPase) was discovered in
1957. It plays an important role in contracting the cardiac muscle, kidney function, and nerve
signaling. The purpose of the sodium-potassium pump is to maintain the proper concentration of
potassium ions K+ and sodium ions Na+ inside and outside of the cell. The Na,K-ATPase pump is a
protein in the cell used to regulate Na+ and K+ gradients across the membrane. As gradients change,
cells can produce electrical signals The outer membrane of each cell is about 70 angstrom in length.
this membrane is made of protein and the cell is surrounded by a double membrane. lipids are found
between the surfaces of both the membranes. the concentration of potassium ion and sodium ion
inside most animal cells are 0.15 M and oh .01 m respectively. The concentration of sodium and
potassium ion in the fluid outside the cells is 0.15 M and 0.004 M respectively thus the large
concentration gradient of sodium and potassium ions requires a sodium pump.
The energy required for the transfer of these ions is obtained by the hydrolysis of atp. active outflow of
sodium ion out of the cell and active entry of potassium ion into the cell by the sodium pump. the
energy obtained from the hydrolysis of one atp molecule is sufficient to drive 3 sodium ions out of the
cell to potassium ion and one hydrogen ion into the cell.
The sodium ion enters the cell along with the glucose molecule,
creating a state of high concentration. therefore it is necessary to do
sodium ion outside the cell. potassium ion are required for the
metabolism of glucose molecules inside the cell. when the
concentration of sodium ion in the inner part of the cell and
potassium ion in the outer part is high then through a
conformational change, the sodium potassium pump moves sodium This is complete nitrogen fixation.
ion out of the cell and potassium ion into The cell. the sodium
potassium adenotriphosphate enzymes of the cell membrane act as
a sodium/ potassium pump.
Na,K-ATPase maintains the proper concentration gradients of Na+ and K+. It is important that Na,K-
ATPase stabilizes the resting membrane potential. If the resting membrane potential of nerve cells is
not stabilized, nerve cells may fire when not needed or may not fire at all. If the Na,K-ATPase stops
working, the concentration gradients of Na+ and K+ on the inside and outside of the cell may not be
correct. This can interrupt cells signals, and cause muscles not to contract, and many other serious
conditions.
 DNA
DNA polymerase is a type of enzyme that is helpful to make
copies of the DNA in the form of Nucleic acid molecules. It is a
group of enzymes required for the synthesis of DNA. It
catalyses the synthesis of DNA during replication. Moreover,
its main function is to duplicate the DNA and divide in cell
division. There are various functionalities of the DNA
polymerase, some of them are – Replication, Repair, and
proofreading. DNA polymerases play an important role in the
synthesis of the DNA and divide it into the cells while the cell
suffers the division by the process of replication.
STRUCTURE & FUNCTION OF MYOGLOBIN SYNTHETIC MODEL COMPLEX OF Cu
Myoclobin is also a pigment similar to hemoglobin it is also produced by hemi and globin. this helical Copper complex Have been designed in order to mimic the properties of hemocyanion. in 1989
is a nuclear protein. it contains only one heme group and 1 polypeptide group. the molecular weight kitajina and coworkers synthesized and characterized Di-Cu(ii) peroxo complex using sterically
of myoglobin its 17,000 stores oxygen in the muscle fibers. oxygen releases it when it is needed the encumburing tri pyra zolyl borate ligand. they found that oxygen binding occurs between two
carrying capacity of myoblobin towards oxygen is greater than That of hemoglobin. hemoglobin and copper center which a side on mode. the compound could be generated at reduced temperature in
myoglobin are attracted to oxygen at high pressure. the pressure in the lungs is high. the pressure of solution by oxygen2 reaction with one or via and acid base reaction of H2O2 with di copper(ii)
oxygen in the muscles is low. here oxygen released by hemoglobin, which is accepted by myoglobin. species.
Myoglobin contains a porphyrin ring with an iron at its center. A proximal histidine group (His-93) is
attached directly to iron, and a distal histidine group (His-64) hovers near the opposite face.[30] The
distal imidazole is not bonded to the iron, but is available to interact with the substrate O 2. This Recently, Simmons and Wilson
interaction encourages the binding of O2, but not carbon monoxide (CO), which still binds about 240× have reported a synthetic
more strongly than O2. copper(I) complex containing
two
De-Oxy myoglobin:- Deoxy Myoglobin contains a imidazole donors that are
five covalently highly cycled iron(iii) complex in capable of binding dioxygen
which the nitrogen item of the porphyrin ring is reversibly in both solid state as
attached at four positions and the nitrogen atom is well as in solution.
attached to the imidazole ligand of the histidine
residue at the 5th position.

Heme group Histadiene

DNA Polymerases Structure :- Most of the structures of the DNA polymerase have a
resembling hand that holds an active site, that site of an enzyme, has two parts – Insertion
site and post-insertion site. Basically, at the insertion site, the nucleotides are appended,
and therefore, after adding, the newly formed base-pair migrates to the site of the post-
insertion.
DNA POLYMERIZATION :- The DNA is the polymer of deoxyribonucleotides and its length
is dependent on the total number of nucleotides present in it.
REPLICATION - Replication is
the main function of DNA
polymerases. It synthesized the
DNA. it maintains and transfers
genetic information from one
generation to another
generation. It works in pairs to
replicate two strands of DNA in
a Tandem. They append DNA at
the 3’-OH group of growing
DNA strands. The DNA grows in
the direction by their
polymerizatn activity based on
5’  3’. The pairing takes place
between Adenine, thymine, and
guanine. The order of pairing is
that the Adenine pairs with
thymine and further, guanine
pairs with cytosine. The main
enzyme responsible for the
replication in prokaryotes is
DNA polymerases While DNA polymerases is the main enzyme for replication in eukaryotes.
REPAIR :- The repair process
is very important for
maintaining integrity in the
genomes because the repair
is a homogeneous function.
Subsequently, it is considered
a continuous process because
it continuously looks if any
error occurs in the genome
because of the DNA
breakdown.
PROOFREADING :- The
process of DNA replication is
not perfect because errors
may occur often. The error
may occur after every 104 to
105 nucleotides added. To
remove the error nucleotides
or the incorrect nucleotides
sequence is very important
for the correct protein
functioning. Proper
functioning is very important
because if not corrected, it
may lead to cancer. The
process of removing the
incorrect pairs by the DNA
polymerases is known as the
exonuclease activity.
Oxy myoglobin:- in oxi myoglobin the oxygen
molecule is coordinated with iron(2) at the sixth
position, causing Fe(2) undergo nitrogen cycling.
the following is spin iron(II) is smaller in size,
causing it to shrink towards the plane Of the
porphyrin ring. this is how myoglobin collects
oxygen in the muscle tissue.
STRUCTURE & FUNCTION OF HEMO-GLOBIN :- Hemoglobin is a type of globular protein present in
red blood cells RBC which transports oxygen in our body through blood.
Hemoglobn structure:- Max Payrutz described the molecular structure of hemoglobin in 1959
hemoglobin is a tetrameric protein. the main type of hemoglobin in adults is made up of two subunits
(α,β = Polypeptide chain)each polypeptide chain is linked to Heme prosthetic group.
Α subunit :- It is made up of alpha polypeptide chain having 141 amino acids residue
β subunit :- it is made up of beta polypeptide chain having 146 amino acid residue
heme group :- it is an iron containing prosthetic group which is attached to each polypeptide chain.
A heme group consists of an iron (Fe) ion held in a
heterocyclic ring, known as a porphyrin. This
porphyrin ring consists of four pyrrole molecules
cyclically linked together (by methine bridges) with
the iron ion bound in the center. The iron ion, which
is the site of oxygen binding, coordinates with the
four nitrogen atoms in the center of the ring, which
all lie in one plane. The heme is bound strongly
(covalently) to the globular protein via the N atoms
of the imidazole ring of F8 histidine residue (also
known as the proximal histidine) below the
porphyrin ring. A sixth position can reversibly bind
oxygen by a coordinate covalent bond, completing
the octahedral group of six ligands.
Oxyhemoglobin is formed during physiological respiration when oxygen binds to the heme
component of the protein hemoglobin in red blood cells. This process occurs in the pulmonary
capillaries adjacent to the alveoli of the lungs. The oxygen then travels through the blood stream to be
dropped off at cells where it is utilized as a terminal electron acceptor in the production of ATP by the
process of oxidative phosphorylation.
Deoxygenated hemoglobin (deoxyhemoglobin) is the form of hemoglobin without the bound oxygen.
The absorption spectra of oxyhemoglobin and deoxyhemoglobin differ. The oxyhemoglobin has
significantly lower absorption of the 660 nm wavelength than deoxyhe hemoglobin, while at 940 nm
its absorption is slightly higher
STRUCTURE & FUNCTION OF HEMOCYANIN
Hemcyanins are protein that transport oxygen throughout the bodies of some invertebrate animals
these metalloproteins contain two copper atoms that reversibly binds a single oxygen molecule they
are second only to hemoglobin in frequency of use as an oxygen transport molecules it contains more
than 100 subunits molecular weight is equal to 4 lakh - 2 crore units.
DE OXY HEMOCYANIN - Both copper are
in +1 state, Diamagnetic nature,
Intermolecular distance between Cu-Cu
is 460 pm., blue colored, Colorless, The
cordination number of each copper is +3,
Distorted triogonal pyramidal geometry.
OXY HEMOCYANIN - Both copper are in
+2 state, Anti ferromagnetic nature,
intramolecular distance b/w Cu-Cu is
360 pm., Cordination number of copper
changes from +5 to 3.
It was found that synthesized
copper(i) complex absorbed 1
mole of oxygen per 2 mole of
copper and this reaction could be
really reversed by gentle heating
and degassing with nitrogen.
SYNTHETIC MODEL COMPLEX OF IRON
Coordination researchers and chemists for many years were unable to develop synthetic iron-
containing oxygen carriers to mimic the natural occurring hemoglobin and myoglobin proteins
since in all the cases Fe (II) complexes were irreversibly oxidised by oxygen to produce μ-oxo Fe
(III)-O-Fe (III) complexes
FeIILn + ½ O2  Ln-1FeIII-O- FeIIIL+2n-1L Dimer/Ireversible complex.
to avoid the formation of μ-oxo Fe complexes in order to successfully fabricate synthetic iron
oxygen carriers and for this purpose they have used following approaches:
(i).. Steric: introduction of the bulky groups on the ligands in such a fashion which sterically
inhibit the formation of Fe-O-Fe dimer (ii) Low temperature: use of low temperature slows down
the rate of reactions leading to dimerization. (iii) Surface attachment: attachment of the iron
complex on the rigid surface prevents dimerization process.
amongst the above methods to synthesize modified porphyrins, steric hindrance has produced
much success.
(i) an imidazole is covalently attached Fe(II) porphyrins . The studies on such tail base
complexes revealed that it leads to the formation of 1:1 adducts only at low temperatures. On the
other hand, the strapped complexes
(ii) and (iii) with a hydrocarbon chain linked over the face of the porphyrin do not undergo
reversible binding with oxygen because of lack of rigidity of the chain, which can be pushed out of
the way. Further, the use of steric constraints has been very nicely demonstrated by Collman and
Baldwin. Collman et al. reported the “picket fence” porphyrins
(iv) Given by collmen and Baldwin, pyrole is substituted by N-alkyl imidazole
(vi) Provides satisfactory steric protection and hence could react reversibly with oxygen in
pyridine.
SYNTHETIC MODEL OF COBOLT
Amongst the several model synthetic dioxygen carriers, cobalt complexes have been the first and
most extensively investigated ones, owing to their structural similarity to those found in the
biological systems . these model complexes have been done using aprotic solvents due to two
reasons
(i).. It was believed that such type of solvents possess the ability to create an environment similar
to that provided by the hydrophobic pocket of the natural oxygen carriers.
(ii) Protonic solvents often lead to the irreversible dioxygen oxidation of the metal complex.
Later, in 1898, these were characterized by Werner and Mylius as ionic and water soluble μ-peroxo
low-spin cobalt (III) complexes (below)
O Co(NH3)5]4+
[(NH3)5Co O [ O - O = 1.47 A ]
t was found that these complexes reacted with oxidizing agents such as cerric ion to generate the
corresponding μ-superoxo cobalt (III) complex described below:

Prokaryotic DNA Polymerases Major type of DNA found in Prokaryotes
1. DNA Polymerases I :- It is the first type of prokaryotic DNA polymerase that is coded
by the polA gene. Its main roles are recombination and repair. It is a Single polypeptide.
Moreover, it has 5’ -> 3’ and 3’ -> 5’ both exonuclease activity. It removes the RNA primer
with the help of 5’ -> 3’ from the lagging strand and it also fills the gap.
2. DNA Polymerases II :- DNA polymerase II is the second type of prokaryotic
polymerase that is coded by the polB gene. The main role of the DNA polymerase II is to
keep backup of the DNA polymerase III and repair. The only exonuclease activity the DNA
polymerase II has is 3’ -> 5’.
3. DNA Polymerases III :- DNA polymerase III is the third type of prokaryotic polymerase
that is coded by the polC gene. It is the main enzyme that is involved in the replication of
E.Coli. The rates of Processivity and polymerization are maximum in this type and it also
has the exonuclease activity of 3’ -> 5’. DNA polymerase III of an E.coli is made of 13 total
subunits that comprise 9 different types of subunits. It is consists of two core domains that
are mainly made up of α, θ, and ℰ subunits.
4. DNA Polymerases IV :- DNA polymerase IV is the fourth type of prokaryotic
polymerase that is coded by the polD gene. Its main action is in the repair of DNA in the
emergency response. When the DNA replication is put at the fork of replication. The other
DNA polymerase types of II, IV, and V are considered as the translesion polymerases.
5. DNA Polymerases V :- It is also involved in the synthesis process of the DNA repair and
response but in the SOS. The DNA polymerase V is made up of the UmuC monomer and
UmuD dimer. Eukaryotic DNA Polymerases
Eukaryotic cells also have many DNA polymerase like the prokaryotic cells but perform
different functions such as mitochondrial DNA replication, nuclear DNA replication, and so
on.The nuclear DNA is mainly initiated by the DNA polymerase types of δ
and α. Certainly, there are at least 15 DNA polymerases identified in human beings.
1. DNA Polymerases δ :- This is considered as the main enzyme for the replication in
eukaryotes with proofreading of 3’ -> 5’ exonuclease activity.
2. DNA Polymerases α :- The main role of this type is to synthesize the primers. The
smaller subunits consist of primase activity while the largest subunit has the
polymerization unit.
3. DNA Polymerases ϵ :- The main function is to Repair the DNA and to remove the
primers from Okazaki fragments from the lagging strand.
4. DNA Polymerases γ :- The main replication enzyme particularly for the Mitochondrial
DNA.
Working of DNA Polymerases
The reaction that shows the working polymerase is the phosphoryl group transfer. The
3 –> OH group of strands that grows acts as the nucleophile and protects from the deoxy
ribo nucleoside tri phosphate by attacking and at the a-phosphate that leads to the
formation of the phosphodiester bond. The reaction is provided below –
(dNMP)n + dNTP → (dNMP)n+1 + PPi
Most of the polymerases require the two Mg ions at the active site and it is mandatory to
take note that DNA polymerase can only append nucleotides at the 3’ end of the growing
strand, because of this the replication always occurs in the 3’ -> 5’ direction.
The replication is considered to be a highly accurate process and the probability of
formation of mutation is high because it can be formed if the change occurs even in a
single nucleotide.
X-ray crystallography of cobalt compounds containing bound dioxygen which shows that the so-
called "inactive" form contains dimeric units [Co(salen)]2 whereas the active forms of Co(salen)
are presumed to contain dimeric units with a more-open lattice packing relative to the inactive
form
STRUCTURE AND FUNCTION OF HEMERYTHRIN
Hemerythrin is a non-heme iron protein used by two phyla of marine invertebrates (sipunculids and
brachiopods) for oxygen transfer and/or storage. It differs from the other oxygen-binding proteins
(hemoglobin and hemocyanin) both in the polypeptide chain and in the metal complex used to
reversibly bind dioxygen.
The two iron atoms in hemerythrin are bound the imidazole rings of five histidine residues and the
carboxylates of an aspartic acid and a glutamic acid. In addition, the complex contains an oxygen atom
bridging between the two iron atoms. In deoxyhemerythrin, the bridge is a hydroxyl group, while in
met- and oxyhemerythrin, the bridge is a mu-oxo atom. In deoxy- and metaquohemerythrin, one of
the iron atoms is bound to six liganding atoms while the other is penta-coordinate. This extra site is
where small molecules such as dioxygen or azide bind to the protein.
In deoxyhemerythrin, the two iron atoms are in the ferrous oxidation state with a bridging hydroxyl
group. As dioxygen is bound to the active site, the hydrogen atom from the hydroxyl bridge moves
over onto the bound ligand, stabilizing the peroxo nature of the bound oxygen molecule. In met-
derivatives of the protein, with and without small molecules bound to the complex, the iron atoms are
both in the ferric oxidation state.
The oxygenated product may be a 1:1 (Co:O2) or a 2:1 (2Co:O2) complex . It is believed that that O2
De oxy & Oxy hemerythrin adds as the sixth ligand to a 5-coordinate complex, or replaces one of the coordinating solvent
molecules; so that the final structure is either a 6 -coordinate 1:1 complex, or a 6-coordinate 2:1
complex.
OXY HEMERYTHRIN One monomeric unit of hemerythrin binds one dioxygen. The dioxygen adds
only to the coordinatively unsaturated ferrous.The dioxygen adds to hemerythrin in an oxidative
manner resulting in the formation of two Fe(III) centers and peroxide (022-).The oxidative addition is
followed by the shifting of proton from the bridged OH to the bound peroxide resulting in the
formation of hydroperoxo (HO2-) group.This proton-transfer result in the formation of a single oxygen
atom (μ-oxo) bridge in oxyhemerythrin.
Most hemerythrin do not bind dioxygen cooperatively. They show tight oxygen binding
HEME PROTEIN AND OXYGEN UPTAKE
The Hemi Group keeps myoglobin and hemoglobin the ability to bind oxygen because of
the presence of iron atom it also contributes to the red color found in muscles and blood
each hammer group contains an iron atom that is able to bind to 1 oxygen molecule each
hemoglobin protein can bind 4 oxygen molecules
Heme protein contain a heme as a prosthetic group. Iron binds to protein either by
incorporation into a protoporphine IX ring or by interaction with other protein ligands.
ferrous and ferric protoporphine IX complex are designated respectively as heme and
hematin. heme containing proteins include those that transport and store oxygen those
involved in the electron transport system and certain enzymes which contain heme as
part of their prosthetic groups.
The Hemi Group consists of Fe+2 ion embedded in a porphyrin ligand the Fe+2 ion has six
coordinates sites. porphyrin ligand takes up only four of the six sites, leaving two free
binding sites on opposite sides of the metal ion. when an oxygen molecule binds to the
iron atom of a heme group though its vacant sixth coordination site, the iron atom
becomes low spin and therefore becomes smaller in radius.
Binding constant of the successive oxygenation reaction of hemoglobin
This is called trigger mechanism the fbo 2 bond in both oxyhemoglobin and oxy myoglobin
is bent as below
CHLOROPHYLL
Chlorophyll is a chlorin pigment, related to the porphyrin containing iron compound
known as heme. At the centre of the ring is a magnesium ion. The side chains vary
somewhat between the different forms of chlorophyll found in different organisms -
chlorophyll a is always present, but chlorophylls b and c also occur in various groups.
Chlorophyll a contains a magnesium ion encased in a large ring structure known as a
chlorin. The chlorin ring is a heterocyclic compound derived from pyrrole. Four nitrogen
atoms from the chlorin surround and bind the magnesium atom. The magnesium centre
uniquely defines the structure as a chlorophyll molecule
Forms of chlorophyll
Chlorophyll consists of two forms, a and b.
a: C55H72O5N4Mg , b: C55H70O6N4Mg
In both cases the magnesium atom is central in the molecule. This green pigment is what
gives green plants their colour. It is involved in photosynthesis by absorbing energy from
visible light. Chlorophyll is a green compound found in leaves and green stems of plants.
Initially, it was assumed that chlorophyll was a single compound but in 1864 Stokes
showed by spectroscopy that chlorophyll was a mixture. If dried leaves are powdered and
digested with ethanol, after concentration of the solvent, 'crystalline' chlorophyll is
obtained, but if ether or aqueous acetone is used instead of ethanol, the product is
'amorphous' chlorophyll.
Function of chlorophyll Due to the green colour of chlorophyll, it has many uses as dyes
and pigments. It is used in colouring soaps, oils, waxes and confectionary. Chlorophyll's
most important use, however, is in nature, in photosynthesis. It is capable of channelling
the energy of sunlight into chemical energy through the process of photosynthesis. In this
process the energy absorbed by chlorophyll transforms carbon dioxide and water into
carbohydrates and oxygen: CO2 + H2O (CH2O) + O2 The chemical energy stored by
photosynthesis in carbohydrates drives biochemical reactions in nearly all living
organisms. In the photosynthetic reaction electrons are transferred from water to carbon
dioxide, that is carbon dioxide is reduced by water. Chlorophyll assists this transfer as
when chlorophyll absorbs light energy, an electron in chlorophyll is excited from a lower
energy state to a higher energy state. In this higher energy state, this electron is more
readily transferred to another molecule. This starts a chain of electron-transfer steps,
which ends with an electron being transferred to carbon dioxide.
TYPES OF CHLOROPHYL
Chlorophyll a - Chlorophyll a is the most widely distributed type of chlorophyll which is
found in all living organisms capable of oxygenic photosynthesis (producing oxygen as a
by-product). It has also been reported in small quantities in some sulfur bacteria
performing anaerobic photosynthesis.
Chlorophyll b is the second most abundant chlorophyll in oxygenic photosynthetic
organisms. These are distinguished from chlorophyll a in the formyl substitution in the C-
7 position of the ring., Chlorophyll b is present as a part of the components of the
peripheral antenna complexes., It is synthesized by the oxidation of the methyl group
present in the chlorophyll a to a formyl group
Chlorophyll c is a form of chlorophyll that acts as an accessory pigment and is less widely
distributed than chlorophyll a and b., These pigments are found in the golden-brown
eukaryotic algae, marine algae, and dinoflagellates.
Chlorophyll d is one of the rarer forms of chlorophylls that are found in some species of
red algae and cyanobacteria., It is mostly found in marine algae, where it acts as an
adaptation to suit algae and photosynthetic organisms found in deep water where not
much light can penetrate.
Chlorophyll e is a rare form of chlorophyll that is found in some golden algae, primarily
two species, Tribonema bombycinum, and Vaucheria hamata., Chlorophyll e is found to be
similar to bacteriochlorophylls that are distributed in cyanobacteria. It acts as an
accessory pigment in different photosynthetic organisms.
Chlorophyll f is the newest form of chlorophyll which was discovered from stromatolites
in 2010., The exact function of chlorophyll f in photosynthesis is not clear yet, but some
evidence of it acting as accessory pigments can be found.
PHOTO SYSTEM
The light absorption processes associated with photosynthesis occur in large protein complexes
present in thylakoid membrane known as photosystems. Photosystems consist of large number of
capturing pigments bound to proteins surrounded by a reaction centre complex. There are two
kinds of photosystems, photosystem I and photosystem II, each capable of capturing light energy.
These photosystems were named in order of their discovery; however photosystem II functions
first in the sequence of steps that make up the light reaction.
overall equation for photosynthesis is 6CO2 + 6H2O - C6H12O6 + 6O2
The plant photosynthetic reactions occur in two stages namely "light reactions" which involve
electron proton transfer processes and dark reactions (often termed as Calvin cycle) which involves
the use of CO 2 for the biosynthesis of carbohydrates. During the light reactions, the solar energy is
converted into ATP and NADPH that are subsequently utilized for driving the light independent
reactions used for the production of carbohydrates. All these vital reactions occur in organelles called
“chloroplasts” containing stacks of thylakoids which appears as a flattened disc. Thylakoid
membranes also contain some integral multi protein complexes known as photosystem I and II which
play an important role in the light-dependent processes of photosynthesis.
Photosystem I Photosystem I is the light-driven plastocyanin-ferredoxin oxidoreductase present in
the thylakoid membranes of cyanobacteria and chloroplasts. It is an extremely efficient biosolar
energy converter, capturing the energy from the sun and converting it into electrical energy through a
light driven charge separation across the membrane. The reaction center of this photosystem contains
chlorophyll a molecules (P700) that absorb light of 700 nm wavelength.
The structure of photosystem I in a cyanobacterium has been provided in above diagram. It is a
homotrimer with each subunit in the trimer containing:
12 different protein molecules bound to:
96 molecules of chlorophyll, 2 molecules of the reaction center chlorophyll P700, 4 accessory
molecules closely associated with them, 90 molecules that serve as antenna pigments
 22 carotenoid molecules,  4 lipid molecules,  3 clusters of Fe4S4,  2 phylloquinones
Working of photosystem
(i) Excitation of electron in Photosystem I: Photoexcited electrons enter photosystem I via
an electron transport chain set in the thylakoid membrane where it waits until the
electron is excited by another photon.
(ii) Chemiosmosis: The energy fall is harnessed to transport hydrogen (H + ) through the
membrane, into the thylakoid lumen to generate ATP.
(iii) Conversion of NADP + to NADPH: The excited electrons in photosystem I
oxidize NADP + to NADPH which will be needed in the Calvin Cycle.
Photosystem II “Photosystem II”- the first link in the photosynthesis chain is a multi-subunit
pigment-protein complex (water-plastoquinone oxidoreductase) embedded in the lipid environment
of the thylakoid membranes of plants, algae and cyanobacteria. At the heart of this photosytem, is a
reaction center (RC) core containing chlorophyll a molecules (P680) that absorbs light at λ max value
of 680 nm. Driven by light, this enzyme catalyzes the chemically and thermodynamically demanding
reaction of water splitting. While doing so, it harnesses solar irradiation to oxidise two molecules of
water to molecular oxygen, liberating electrons which provide the reducing equivalents required for
the conversion of CO 2 into the organic molecules of life. Photosystem II is also a complex assembly of
more than 20 different protein molecules bound to:
 50 or more chlorophyll a molecules, 2 molecules of the reaction center chlorophyll P 680,
2 accessory molecules close to them, 2 molecules of pheophytin (chlorophyll without the Mg ++ ),
the remaining molecules of chlorophyll a serve as antenna pigments.,  Some half dozen carotenoid
molecules,also serve as antenna pigments.,  2 molecules of plastoquinone
Working of photosystem II
 Light absorption:Absorption of light energy by photosystem II (P680).
 Electron capture: Excitation of electron from a low energy state to a high energy state. The electron
travels down the electron transport system (ETS). Along the way, more H+ is pumped into the
thylakoid compartment.
 Splitting ofwater and releasing oxygen: Meanwhile the photosystem's lost electron is
replenished by photolysis, or the splitting of H 2 O to form H + and O2 (note: the H+ is kept
inside the thylakoid membrane). The O 2 resulting is the source of all oxygen in our atmosphere
How do photosystems I and II work together?
The photosystems work through resonance effects shown below. When light is absorbed by
photosystem II, the electrons present in the reaction-center get excited which are eventually trapped
by the primary electron acceptors. The hole thus created in the reaction centre by the departure of
photo-excited electron is replenished by the electrons extracted from water through a cluster of four
manganese atoms in photosystem II. In this process, oxygen is released when four electrons have
been removed from two molecules of water. Further, the light energized electrons travel to
photosystem I through the cytochrome b6f complex (an enzyme found in the thylakoid membrane
that catalyze the transfer of electrons from plastoquinol to plastocyanin) via an electron transport
chain. During this process, often termed as chemiosmosis, transfer of H+ ions takes place across the
thylakoid membrane via plastoquinone which creates a gradient within the chloroplast that is used
for the production of ATP molecules. When electron enters photosystem I, it waits until the electron is
excited by another photon. The electron thus excited is captured by another electron acceptor which
flow down a chain of electron carriers to NADP + to reduce it to NADPH that enters the Calvin cycle.
2H2O + 2NADP+ + 8 photons -> O2 + 2 NADPH + 2H+
STORAGE OF GLUCOSE
Glucose is the most simple sugar of carbohydrates. When more complex carbohydrates such as
polysaccharides and disaccharides are broken down in the stomach, they break down into the
monosaccharide glucose. Carbohydrates serve as the primary energy source for working muscles,
help brain and nervous system functioning and help the body use fat more efficiently.
Carbohydrates are present in most of the food we intake such as bread, fruits, vegetables, meats
and so on. Any food that contains sugar has carbohydrates. And, most foods are converted to
sugars when they are digested Once carbohydrates are absorbed from food, they are carried to the
liver where its processing occurs. Some glucose is sent to the bloodstream for its availability to
various tissues while the rest is stored for later requirement of energy. It is stored in two types as
muscle glycogen and as liver glycogen.
Function of Glucose
Two most important function of glucose are-
1. The primary function of glucose is to provide energy for physiological processes such as
respiration, muscle contraction and relaxation, heart rhythm and the regulation of body
temperature.
2. The energy needed by the brain, neurons and developing red blood cells can only be acquire
from glucose.
Muscle stores approximately 1200 kcal or 5020 kJ amount of glycogen. This glycogen is readily
converted into glucose 6-phosphate for entry into glycolysis. Muscle cannot export glucose because
it lacks glucose -6-phosphatase. Furthermore, although muscle can synthesize glycogen from
glucose, it does not participate in gluconeogenesis because it lacks the required enzymatic
machinery. Consequently, muscle carbohydrate metabolism serves only muscle. In actively
contracting skeletal muscle, much of the pyruvate formed is reduced to lactate, some of which
flows to the liver, where it is converted into glucose.
Liver The liver maintains the
proper levels of circulating fuels
for use by various tissues such as
brain and muscles. One of the
major functions of liver is to act
as a blood glucose ‘buffer’. It does
so by taking up and releasing
glucose in response to hormones
and to the concentration of
glucose itself. After a
carbohydrate containing meal,
when the blood glucose
concentration reaches ≈ 6mM,
the liver
takes up glucose by converting it
to glucose-6-phosphate. The liver
can produce glucose for release
into the blood by breaking down
its store of glycogen and by
carrying out gluconeogenesis
Storage of glucose in plants
The storage form of glucose in plants is starch. Starch is a polysaccharide of the monomer
glucose as can be seen in the figure 6. The leaves of a plant make sugar during the process of
photosynthesis using chlorophyll present in them. Photosynthesis occurs in light (photo = light)
when the sun is shining as the energy from the sunlight is used to make energy for the plant. So,
when plants are making sugar, they store some of it as starch. When the simple sugars need to be
retrieved for use, the starch is broken down into its smaller components.
Glycogen
Glycogen (in animals, fungi and bacteria) and starch (in plants) can function to stockpile glucose
for later metabolic use. In animals, a constant supply of glucose is essential for tissues such as the
brain and red blood cells, which depend almost entirely on glucose as an energy source. The
mobilization of glucose from glycogen stores, primarily in the liver, provides a constant supply of
glucose (≈ 5 mM in blood) to all tissues. When glucose is plentiful, such as immediately after a
meal, glycogen synthesis accelerates. Yet the liver’s capacity to store glycogen is sufficient to
supply the brain with glucose for about half a day. Under fasting conditions, most of the the
body’s glucose needs are met by gluconeogenesis from noncarbohydrate precursors such as
amino acids.
Structure of Glycogen
Glycogen is a polymer of α(1→4)-linked D-glucose with α(1→6)-linked branches every 8-14
residues (Figure 7). Glycogen occurs as intracellular granules of 100 to 400 Å diameter
spheroidal molecules that each contain up to 120,000 glucose units. The granules are especially
prominent in the cells that make the greatest use of glycogen: muscle (up to 1-2% glycogen by
weight) and liver cells (up to 10% glycogen by weight). Glycogen granules also contain the
enzymes that catalyze glycogen synthesis and degradation as well as many of the proteins that
regulate these processes.
In this structure of two outer branches of a glycogen molecule, the residues at the nonreducing
ends are shown in red and residue that starts a branch is shown in green. The rest of the glycogen
molecule is represented by R