The process of synthesizing ATP from ADP

and Pi coupled with the electron transport

chain is known as oxidative phosphorylation.

The complex 5 of the inner mitochondrial membrane is the site of oxidative phosphorylation.

Several hypothesis have been put forward to explain the process of oxidative phosphorylation.

The most important among them namely,

CHEMICAL COUPLING CHEMIOSMOTIC HYPOTHESIS

This hypothesis put forward by Edward Slater in 1953. According to chemical coupling hypothesis during the course of electron transfer in respiratory chain, a series of phosphorylated high energy intermediates are first produced which are utilised for the synthesis of ATP. However this hypothesis lacks experimental evidence so this have not been successful.

This mechanism proposed by Peter Mitchell in 1961, now widely accepted. It explain how the transport of electrons

through the respiratory chain is effectively

utilised to produce ATP from ADP and Pi.

The inner mitochondrial membrane is impermeable to proton H+ and hydroxyl ions OH_.

The transport of electron through ETC is

coupled with the translocation of protons across the inner mitochondrial membrane from matrix to inner membrane space.

The pumping of protons result in an eletrochemical or proton gradient. This is due to the accumulation of more H+ ions on the outer side of inner mitochondrial membrane than the inner side.

the proton gradient developed due to the

electron flow in the respiratory chain is

sufficient to result in the synthesis of ATP

from ADP and Pi.

ATP synthase , present in the complex 5 utilises

the proton gradient for the synthesis of ATP.

The enzyme is also known as ATP ase since it can hydrolyse ATP to ADP and Pi. ATP synthase is a complex enzyme and consist of two functional subunit namely, F0 and F1.

Paul Boyer in 1964 proposed that a conformational change in the mitochondrial membrane protein leads to the synthesis of ATP. The enzyme ATP synthase is F0F1 complex of complex 5. The F0 subcomplex is composed of channel protein C subunit to which F1 ATP synthase is attached.

F1 ATP synthase consist of a central gamma subunit surrounded by alternating alpha and beta subunit.

In response to the proton flux , the gamma

subunit physically rotates.

This induces conformational change in the

beta subunit that finally leads to the release of ATP.

According to the binding change mechanism, the three beta subunit of F1 ATP synthase adopt different conformation.

One subunit has open (o) conformation, the

second has loose (L) conformationwhile the third one has tight (T) conformation. By an unknown mechanism, protons induce the rotation of gamma subunit, which inturn induces

conformational changes in beta subunit.

The substrate ADP and Pi bind to subunit in L conformation. The L site changes to T conformation and this leads to the synthesis of ATP. the O site changes to L conformation which binds to ADP and Pi. The T site changes to O conformation and release of ATP. This cycle is repeated and ATP are generated for each revolution. Thus ATP synthase is worlds smallest molecular motor.

It is estimated that about 100 polypeptides are

required for oxidative phosphorylation.

Of these 13 are coded by mt DNA are synthesised in the mitochondria , while the rest are produced in the cytosol. Mitochondrial DNA is maternally inherited and is

about 10 times more susceptible to mutation

than nuclear DNA.

Lebers heriditary optic neuropathy is an

example for mutation in mt DNA is characterised by loss of bilateral vision due to neuroretinal degeneration.

INHIBITORS OF ETC

The synthesis of ATP is dependent on

electron transport . Hence all the site specific inhibitor of ETC also inhibit ATP formation.

The 3 possible site of actions for the inhibitors of ETC are, NADH and coenzyme Q: Fish poison rotenone, barbituate drug amytol and antibiotic piercidinA inhibit this site. Between cytochrome b and c1: Antimycin A , an antibiotic British antilewisite an antidote used against war gas are the two important inhibitors of the site. Inhibitors of cytochrome oxidase: CO, cyanide,effectively inhibit cytochrome oxidase

The inner mitochondrial membrane is

impermeable to NADH.

Therefore NADH produced in the cytosol cannot directly enter the mitochondria. Two pathways namely:

1,Glycerol phosphate shuttle 2,malate aspartate shuttle do this job.

Cytosolic glyceol - 3- po4 dehydrogenase oxidises NADH to NAD+. The reducing equivalents are transported through glycerol -3 - po4 in to the mitochondria. Glycerol - 3- po4 dehydrogenase present on outer surface of inner mitochondrial membrane reduces FAD to FADH2 oxidised via ETC to generate 2 ATP.

In the cytosol oxaloacetate acccepts NADH and become malate. Malate then enters mitochondria where it is

oxidised by mitochondrial malate

dehydrogenase.

In this reaction NADH gets oxidised via ETC and 3 ATP are produced.

Liver , heart, spleen, kidney utilize malate aspartate shuttle and yield 3 ATP per mole of NADH.

Brain , skeletal muscle and other tissue

employ glycerol po4 shuttle and liberate 2 ATP from NADH.

Grouped in to 4 categories

1, Oxidase 2, Dehydrogenase

3, Hydroperoxidase
4, Oxygenase

OXIDASE: Enzyme catalyse the elimination of H from substrate is accepted by O to form water. Eg:cytochrome oxidase DEHYDROGENASE: As the name indicate these enzyme cannot utilise O. They catalyse the reversible transfer of H from one substrate to other, thus bring about oxidation reduction reaction. Eg: FMN dependent dehydrogenase

HYPERPEROXIDASES: There is a constant production of H2O2 in the reaction catalysed by aerobic dehydrogenase.The

harmful effect of H2O2 are prevented by

hydroperoxidase. OXYGENASES Group of enzyme catalyses the direct incorporation of O in to the substrate molecule.

Inorder to maintain living process, all organisms must obtain supplies of free energy from their environment. Autotrophic organisms utilize simple exergonic processes, eg:the energy of sunlight (green plants). On the other hand, heterotrophic organisms ATP plays a central role in the transferance of free energy from the exergonic to the endergonic process.

The high energy compounds when hydrolysed will release a large quantities of energy. The high energy bond in compounds is usually indicated by a squiggle bond (~). Examples of high energy compounds are: 1. Nucleotides ATP, GTP, UTP, UDP GLUCOSE 2. Creatine po4 3. Arginine po4 4. Phosphoenol pyruvate 5. Amino acyl AMP 6. Acetyl co A

ATP is the universal currency of energy within the

living cells.

The hydrolysis of ATP to ADP under standard conditions release -30.5 kJ/mol or -7.3kcal/mol. An average person at rest consumes and regenerates ATP at a rate of approximately 3 molecules per second, ie about 1.5 kg/day.

CP provides a high energy reservoir of ATP to

regenerate ATP rapidly by the Lohmanns reaction

catalysed by creatine kinase. ATP + creatine 43.1KJ/mol The reaction is mitochondrial and of special phosphocreatine +ADP

significance in the myocardium which has a high

energy requirement, about 6kg of ATP per day.