1. You design lecture a.Discuss the research that “proved” Rotational Catalysis b.Review Metabolism and Electron Transport Chain from previous two classes c.Explain how Glycolysis is like eating Pizza! d.Talk about Fermentation as an option in some cells e.Discuss Krebs Cycle and the path inside the mitochondria like the events in the Biovisions movie

2. Masasuke Yoshida “Ummm… check this out.”

6. Hiroyasu Itoh –designed this approach (Nature 2004) “Reporter enzyme” emits light when detects new ATP

8. Analyze the data. Given that the gray bars represent the basal/background level of ATP in the experiment, explain what you believe would be (i) predictions v. (ii) observations

9. Figure 1 Itoh et al, Nature 2004 9

10. You design lecture

13. Proteins Carbohydrates Amino acids Sugars Fats GlycerolFatty acids Glycolysis Glucose Glyceraldehyde-3- Pyruvate P NH 3 Acetyl CoA Citric acid cycle Oxidative phosphorylation

14. Enzyme 1 Enzyme 2 Enzyme 3 Start of pathway X Presence of product inhibits enzyme 1 Homeostasis: What is negative feedback? Inter- mediate Inter- mediate Product

15. Glucose Glycolysis Fructose-6-phosphate Phosphofructokinase Fructose-1,6-bisphosphate Inhibits AMP Stimulates Inhibits Pyruvate Citrate Acetyl CoA Citric acid cycle Oxidative phosphorylation ATP + – –

16. The electron transport chain occurs in the inner membrane of the mitochondrion (in regions called “cristae”) ELECTRON TRANSPORT CHAIN H+H+ NAD + FAD Complex IComplex IIComplex IIIComplex IV Cyt c FADH 2 NADH Inner membrane O2O2 Q H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H2O H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ F o unit Stalk F 1 unit ATP ADP+PiPi H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ Q ATP

19. You design lecture

20. ELECTRON TRANSPORT AND OXIDATIVE PHOSPHORYLATION FERMENTATION Cellular respiration KREBS CYCLE Glucose Pyruvate GLYCOLYSIS If electron acceptor (such as oxygen) is present If electron acceptor (such as oxygen) is NOT present

21. ADP GLYCOLYSIS All 10 reactions of glycolysis occur in cytosol Glucose- 6-phosphate What goes in: What comes out: 2 Fructose- 6-phosphate Fructose- 1,6-bisphosphate ATP Glycolysis begins with an energy- investment phase of 2 ATP ADP ATP 1345 Glucose

22. PIP Think of the beginning of “Glyco-lysis” as eating a pizza

23. Think Glucose = Pizza The common ‘hexo-pyranose’ form of Pizza

24. Meet PIP (Phosphorylate-Isomerize-Phosphorylate) Is that really the best way to hold on to your pizza?

25. Aldolase = Fructose 1-6 Bisphosphate -> DHAP (ick) and G-3-P (tasty!)

26. DHAP (ick) G-3-P (tasty!) Isomerase ->

27. G-3-P (tasty!)

28. ADP GLYCOLYSIS Glucose- 6-phosphate What goes in: What comes out: 2 Fructose- 6-phosphate Fructose- 1,6-bisphosphate ATP Energy-investment phase ADP ATP 1345 Glucose Hexokinase (capture) PFK*phosphoglucoisomerase [ATP] high PPI ??????

29. ATP at regulatory site Fructose- 1,6- bisphosphate at active site ATP at active site PFK is highly regulated

30. ADP GLYCOLYSIS Glucose- 6-phosphate What goes in: What comes out: 2 Fructose- 6-phosphate Fructose- 1,6-bisphosphate ATP Energy-investment phase ADP ATP 1345 Glucose hexokinasePFK*phosphoglucoisomerase aldolase isomerase DHAP [ATP] high PPI G-3-P inhibits Glyceraldehyde-3-phosphate (G3P)-dehydrogenase [ADP] [AMP] stimulates

31. Phosphoglycero(PG)-kinase Glyceraldehyde-3-phosphate(G3P)-dehydrogenase PIP

32. ADP GLYCOLYSIS All 10 reactions of glycolysis occur in cytosol Glucose- 6-phosphate What goes in: What comes out: 2 Fructose- 6-phosphate Fructose- 1,6-bisphosphate ATP Glycolysis begins with an energy- investment phase of 2 ATP ADP ATP 1345 Glucose G-3-P Glyceraldehyde-3-phosphate(G3P)-dehydrogenase G-3-P

33. During the energy payoff phase, 4 ATP are produced for a net gain of 2 ATP 768910 2 2 NAD + 2 NADH 2 ADP 2 ATP Pyruvate 2 ATP 2 ADP 2 2 22 The “2” indicates that glucose has been split into two 3-carbon sugars Phosphoglycero(PG)-kinase 3-PGA1,3-BisPGA G-3-P-dehydrogenase 2 G-3-P Energy payoff phase pyruvate kinase

34. You design lecture

41. SUMMARY OF GLUCOSE OXIDATION Mitochondrion Glucose 2 NADH 2 Pyruvate Cytoplasm 2 ATP Where do we go next?

42. Pyruvate Dehydrogenase Multienzyme Complex

45. SUMMARY OF GLUCOSE OXIDATION Mitochondrion Glucose 2 NADH 2 Pyruvate Cytoplasm 2 ATP If you have O2, enter the Matrix

46. SUMMARY OF GLUCOSE OXIDATION Mitochondrion 2 Acetyl CoAGlucose 2 NADH 2 CO 2 2 Pyruvate Cytoplasm 2 ATP 2 NADH Where do we go next?

47. SUMMARY OF GLUCOSE OXIDATION Mitochondrion 2 Acetyl CoAGlucose 2 NADH 6 NADH 2 FADH 2 2 ATP 2 CO 2 4 CO 2 2 Pyruvate KREBS CYCLE Cytoplasm 2 ATP 2 NADH

48. Is this what the Citric Acid Cycle looks like?

49. Is this what the Citric Acid Cycle really looks like?

50. What about this?

51. Actually…

53. You design lecture