1. Week 8 Aerobic cellular respiration continued and Membrane structure

2. Review Glycolysis –products include: 2 molecules of pyruvate 2 ATPs via substrate level phosphorylation 2 NADH (reduced NAD + ) –NAD + is required to keep glycolysis going. How is NAD + regenerated from NADH? –What is the fate of pyruvate?

3. Kreb’s cycle Syn. Tricarboxylic acid cycle (TCA) and Citric acid cycle Occurs in matrix of mitochondria Requires: –pyruvate –oxaloacetate from Kreb’s cycle –indirectly requires oxygen

4. Kreb’s cycle Pyruvate enters mitochondria Pyruvate + CoASH --> acetyl-CoA + NADH + CO 2 –enzyme: pyruvate dehydrogenase complex –hydrolysis of citrate-CoA intermediate releases lots of energy that is used to drive the above reaction to the right. –See panel 4-2panel 4-2

6. Kreb’s cycle See panel 4-2panel 4-2 Key points: –you need to be able to account for the number of carbons in each intermediate. Important intermediate compounds –citrate –  -ketoglutarate –oxaloacetate

8. Kreb’s cycle Summary of products starting with acetyl CoA –2 CO 2 released - complete oxidation of acetate –1 ATP (GTP) –3 NADH –1 FADH 2 What is the fate of NADH and FADH 2 ?

9. So, What is the fate of NADH and FADH 2 ? Like glycolysis, without NAD + and FAD, the Kreb’s cycle would shut down. They are required cofactors for enzyme activity. NADH and FADH 2 pass their high energy electrons to the electron transport chain embedded in the inner membrane of the mitochondrion.

10. Electron transport chain The electrons move down the electron transport chain (see figure 13-9) and generate a proton motive force. Oxygen, indirectly required by the Kreb’s cycle, is the terminal electron acceptor of the electron transport chain. The proton motive force is the driving force behind oxidative phosphorylation via ATP synthase.

11. Yield of ATP ATP yields: –Aerobic cellular respiration (glycolysis, TCA cycle, and electron transport chain activities) yields approximately 30 ATP from glucose –glycolysis alone yields only 2 ATPs per glucose

12. Anaerobic conditions Exercise physiology –strenuous exercise causes muscles to be depleted of oxygen. What happens? –Lactic acid builds up -burning sensation in muscle. Why? –Because the oxygen level has dropped, NADH and FADH 2 are not being oxidized back to NAD+ and FAD.

13. Anaerobic cellular physiology Oxygen is depleted through normal cellular respiration during strenuous exercise. To oxidize NADH to NAD + needed for glycolysis, muscles use pyruvate as a terminal electron acceptor in what is called lactic acid fermentation. Pyruvate + NADH --> lactic acid + NAD + NAD + can return to function in glycolysis.

14. Alcohol fermentation Yeast can grow either aerobically using aerobic respiration or anaerobically using alcohol fermentation. Under anaerobic conditions, pyruvate is oxidized to acetaldehyde + CO 2 Acetaldehyde + NADH --> ethanol + NAD + NAD + can function in glycolysis again.

15. Comparison of aerobic respiration and alcohol fermentation Alcohol fermentation yields ATP via substrate level phosphorylation only during glycolysis. ONLY 2 ATPs per glucose yielded Aerobic respiration yields ATP via both substrate level phosphorylation and oxidative phosphorylation. Up to 30+ ATPs yielded per glucose!

16. Anabolic pathways A number of intermediates of both glycolysis and Kreb’s cycle are used as substrates for synthesis of nucleotides, amino acids, lipids etc. See Figure 4-18.4-18

18. Poisons of respiration Rotenone: –blocks NADH dehydrogenase (complex I). NADH cannot transfer its high energy electrons to the electron transport chain in the mitochondria. Therefore the cells cannot generate a proton motive force and no ATP can be made by oxidative phosphorylation. The cells sense death!

19. Poisons cont Dinitrophenol - used as a diet supplement in the 1960s –Dinitrophenol is called an uncoupler of oxidative phosphorylation. It makes the inner membrane of mitochondria permeable to protons and diffuses the proton gradient. Electrons move through the electron transport chain and try to make a proton gradient. But the membrane is permeable to protons which disrupts the proton motive force and very little ATP is made by ATP synthase. Instead the energy of the electrons moving down the electron transport chain is released as heat.

20. Poisons cont Arsenate –chemically similar to inorganic phosphate. –Competes where inorganic phosphate is used to make ATP via substrate level phosphorylation, e.g., glycolysis and Kreb’s cycle

21. Poisons cont Cyanide –interferes with complex III which is the complex that reduces oxygen to water. –If this is blocked, than the electron transport chain would be saturated with electrons. Therefore NADH and FADH 2 would not be able to be oxidized to NAD + and FAD respectively. Then glycolysis and Kreb’s cycle would be negatively effected.

22. Hibernating bears Bears have lots of BAT- brown adipose tissue (fat cells rich with mitochondria) During hibernation the fat of these cells is metabolized to yield heat. –How? Uncouple ATP synthesis from proton motive force. Use the energy of the electrons to generate heat instead of ATP

23. Membranes

24. Function: –selective barrier between cytoplasm and its contents and outside the cell. –Composed of phospholipid, other lipid material and proteins

25. Phospholipids Composed of a phosphate head, glycerol, and two fatty acids. (see Fig. 11-6)Fig. 11-6 Amphipathic molecule –means that there is a hydrophilic and hydrophobic region - the phosphate head is hydrophilic and the fatty acid tails are hydrophobic

27. Liposomes See Fig. 11-13Fig. 11-13 Formed from phospholipids Lipid bilayer with an aqueous interior and exterior environments. Energetically favorable structure due to the amphipathic nature of the phospholipids.

29. membranes Plasma membrane –circa 50 nm thick lipid bilayer –separates inside from outside environment of a cell –Bacteria have a single plasma membrane –Eucaryotes have a plasma membrane and numerous internal membranes and membrane bound organelles.

30. Lipid bilayer See Fig. 11-4Fig. 11-4 Types of lipids –See Fig. 11-7Fig. 11-7 –phospholipids –sterols –glycolipids

33. Lipid bilayer Fluid bilayer lipids float freely in their plane of the bilayer –rarely flip to otherside of bilayer –enzyme called flipase carries out flips. Fluidity function of composition of fatty acids in phospholipids. –Short, unsaturated fatty acids increase fluidity –long, saturated fatty acids decrease fluidity

34. Saturated vs unsaturated fatty acids CH3 - CH2 - CH2 - CH2 - CH2 - CH2 - COOH = saturated fatty acid vs CH3 - CH2 - CH2 - CH = CH - CH2 - COOH = unsaturated fatty acid

35. Adaptation of membrane phospholipids As the temperature goes up, some yeast and bacteria can reduce the number of double bonds and increase the length of the fatty acids of their phospholipids. Yeast can also include cholesterol in their membranes to reduce the fluidity as the temperature goes up.

36. Lipid bilayer See Fig. 11-17Fig. 11-17 Asymetrical in composition of phospholipids –e.g., glycolipids found on non-cytosolic side of membrane –inositol phospholipids found on cytosolic side of membrane