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LET'S TALK LIFE-SCIENCE BIOCHEMISTRY

Suraj Prakash Sharma | Ekta Chotia

MECHANISMS OF OXIDATIVE PHOSPHORYLATION
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16.3.1.   Heat Production

The proton gradient generated by electron transport can be used to generate heat. Heat in human infants and some hibernating animals is produced by brown fat. For example, human infants, other mammals born hairless and some hibernating animals have a special type of brown fat in the neck and upper back.

The brown fat is so named because it contains profuse mitochondria which contain red-brown cytochromes. The brown fat mitochondria has special proton channels in their inner membrane proton to flow back to matrix. These specialized  brown-fat mitochondria do not usually produce ATP, rather they dissipate the free energy of electron transport as heat in order to maintain the body temperature of the young ones.

This is because the brown-fat mitochondria have special proton pores in their inner mitochondrial membrane. The proton pores allow the protons, pumped on by electron transport, to flow back into the matrix, rather than through the F0F1 ATPase or ATP synthetase. Consequently, the free energy of electron transport is diverted from ATP synthesis into heat production.

16.3.2.   Transport of Ca+2

The electron transport energy is also used to transport Ca+2 from the cytosol into the matrix of animal mitochondria so as to maintain the concentration of Ca+2 low inside the cytoplasm. High Ca+2 concentrations promote many cell functions such as muscle concentration, glycogen breakdown and the oxidation of pyruvate.

16.3.3.   Bacterial Movement

In bacteria the rotation of  flagella is controlled by the proton gradient generated across the membrane.

16.3.4.   Other Function

  1. Transfer for electrons from NADH to NADPH is also powered by the proton gradient.
  2. The energy generated during electron transport also help in transport of some amino acids and sugars.

16.4.      Uncouplers of Oxidative Phosphorylation

The transport of electrons is coupled with the synthesis of ATP. Uncoupling agents dissociate (or ‘uncouple’) these two process.  This means that the electron transport continues to function, leading to oxygen consumption but phosphorylation of ADP is inhibited, and the energy release in the form of Heat. This can be done by increasing the permeability of the inner membrane to H+.

The chemical nature of uncoupler is lipophilic and they bind H+ from perimitochondrial space and transport them to matrix. As the uncouplers bind and carry protons, they are also called protonophores. In Mitchell’s hypothesis, uncouplers are agents that are capable of destroying the vectorial, anisotropic structure of the membrane, leading to elimination of the pH gradient.

16.4.1.   Some uncouplers are :

1.            2, 4-dinitrophenol (DNP).           

It does not effect the substrate-level phosphorylation of glycolysis. At pH 7.0 DNP  exists mainly as the anion which is not soluble in the lipids. In its protonated form, it is lipid-soluble and hence can pass through inner membrane, carrying a proton and transport the proton to other side. The proton (H+) carried by DNP is discharged on the other side of the membrane. The phenolate ion then diffuses back towards the cytosol side, where it picks up a proton to repeat the process. In this way, uncouplers prevent formation of H+ gradient across the membrane. Dinitrophenol also stimulates the activity of the enzyme ATPase., which is normally inactive as a hydrolytic enzyme in mitochondria. Actually, ATP is never formed in the presence of DNP, since the high-energy intermediate is attacked i.e., it acts prior to the step of ATP synthesis.

2.            Dicoumarol

It has an action identical to that of 2,4-dinitrophenol. Dicoumarol is also an antagonist of vitamin K function.

3.            m-chlorocarbonyl cyanide phenylhydrazone (CCCP)

Its action is also similar to that of 2, 4-dinitrophenol but it is more active than the DNP.

16.5.      Energy Balance

ATP obtained from a complete catabolism of glucose

1.  From Glycolysis (in cytoplasm)

     For each glucose 2 ATP’s used                 -2

    4 ATP’s formed                                            +4

The NADH produce in glycolysis can yield either 2.5 ATP or 1.5 ATP depending upon the shuttle system.

     2NADH (Glycolysis)                                    5 or 3

2.  2 Pyruvate → 2 acetyl-CoA

     2 NADH molecules formed (2.5 ATP)       +5

(This NADH is already in the mitochondria and no transport is necessary.)

3.  Citric Acid Cycle (and Electron transport chain)

From each acetyl-CoA we get 3 NADH, 1 FADH2 and 1 ATP. Two acetyl-CoA enter the cycle (if we started with 1 glucose).

      6 NADH (2.5 ATP)                                     +15

      2 FADH2 (1.5 ATP)                                    +3

      2 ATP                                                          +2

      Total  (for two)                                         30/32

16.6.      Shuttle system :

The glycerophosphate shuttle. The electrons of cytosolic NADH are transported to the mitochondrial electron-transport chain in three steps : (1) Cytosolic oxidation of NADH by dihydroxyacetone phosphate catalyzed by glycerol-3-phosphate dehydrogenase.

This enzyme is present in cytosolic (2) Oxidation of glycerol-3-phosphate by flavoprotein dehydrogenase with the reduction of FAD to FADH2. (3) Reoxidation of FADH2 with the passage of electrons into the electron-transport chain.

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