BIOSCIENCE MODULE TASK I

BIOLOGICAL OXIDATION

Questions: 1. Mitochondrion is an organelle of the cell that is often mentioned as “the centre of cell energy” through the phosphorylation oxidation process to produce ATP. Please explain (accompanied with picture) how the phosphorylation oxidation process occurs in the mitochondria to produce ATP! 2. There are many toxic materials that inhibit the phosphorylation oxidation reaction. Please mention and explain the mechanism of that inhibitor! 3. The hydroperoxidase enzyme plays an important role in protecting the body from the free radical damage from the reactive oxygen species. Explain the mechanism of the enzyme! 4. The ATP synthase enzyme plays an important role in the process of the ATP formation as the source of cell energy. Please explain how ATP synthase enzyme produces ATP!

By Theo Dapamede. 2009.

Answers: 1. In order for oxidative phosphorylation to proceed, two principal conditions must be met. First, the inner mitochondrial membrane must be physically intact so that protons can only re-enter the mitochondrion by a process coupled to ATP synthesis. Second, a high concentration of protons must be developed on the outside of the inner membrane.1 Each of the respiratory chain complexes I, III, and IV act as a proton pump. The inner membrane of the mitochondria is impermeable to small molecules and ions including protons. Protons that are pumped by the proton pumps accumulate outside the inner membrane, creating an electrochemical potential difference across the membrane, which consists of a chemical potential (difference in pH) and an electrical potential. This electrochemical potential difference is then used to drive a membrane located ATP synthase which in the presence of Pi and ADP forms ATP.

2. Inhibitor of the oxidative phosphorylation reaction: a. Rotenone: inhibits electron transport from complex I b. Amytal: inhibits electron transport from complex I c. Antimycin A: inhibits electron transport from complex III d. Cyanide: inhibits electron transport from complex IV e. Carbon Monoxide: inhibits electron transport from complex IV f. Azide: inhibits electron transport from complex IV g. 2,4-dinitrophenol: acts as an uncoupling agent at the trans-membrane H+ carrier h. Pentachlorophenol: acts as an uncoupling agent at the trans-membrane H+ carrier
1 Michael King, PhD. 2009. http://themedicalbiochemistrypage.org/oxidativephosphorylation.html.

By Theo Dapamede. 2009.

i. Oligomycin: inhibits ATP synthase. 3. The enzyme hydroperoxidase uses hydrogen peroxide or organic peroxide as the substrate. Hydrogen peroxide formed from the reaction of superoxide anion free radical removal by superoxide dismutase (SOD) will be used as substrate by hydroperoxidase to undergo a reduction-oxidation reaction and form H2O.
4. The ATP synthase consists of an F0 sub-complex, which is a disk of “C” protein

subunits. Attached to the F0 is a γ-subunit in the form of a “bent axle”. Protons passing through F0 sub-complex cause it and the attached γ-subunit to rotate. The γsubunit fits inside the F1 sub-complex of three α- and three β-subunits that are fixed to the membrane and do not rotate. ADP and Pi are taken up sequentially by the βsubunits to form ATP and are expelled as the rotating γ subunit “squeezes” each βsubunit in turn. Thus, three ATP molecules are generated per rotation.

By Theo Dapamede. 2009.

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