1. Citric Acid Cycle 1 C483 Spring 2013

2. 1. The net effect of the eight steps of the citric acid cycle is to A) completely oxidize an acetyl group to carbon dioxide. B) convert pyruvate to Acetyl CoA. C) produce a citrate molecule D) produce 8 ATP for every pass through the cycle. E) More than one of the above 2. The order of prosthetic groups as they act in the three proteins of the PDH(pyruvate dehydrogenase) complex is: A) FAD → thiamine pyrophosphate → NAD+ B) FAD → thiamine pyrophosphate → dihydrolipoamide C) thiamine pyrophosphate → dihydrolipoamide → FAD D) NAD+ → FAD → dihydrolipoamide 3. Which might you expect to have a higher than normal blood concentration in an individual with thiamine deficiency? A) Isocitrate. B) Pyruvate. C) Oxaloacetate. D) Acetyl CoA.

3. 4. About how many total ATP equivalents are generated by the complete oxidation of one molecule of acetyl CoA? A) 1.5 B) 2.5 C) 3 D) 10 E) 30 5. Which product of the citric acid cycle produces the most ATP equivalents? A) NADH. B) QH2. C) GTP. D) CO2.

4. Overview Compartmentalization – Glycolysis: Cytosol – Citric Acid Cycle: mitochondria

5. Overview Glycolysis Pyruvate dehydrogenase complex – Commitment of carbon away from carbohydrates Citric acid cycle

6. Pyruvate Dehydrogenase Complex Three distinct enzymes—in a massive complex Five chemical steps What cofactors needed?

7. Pyruvate Dehydrogenase (E 1 ) TPP cofactor: draw mechanism of decarboxylation

8. Dihydrolipoamide Acyltransferase (E 2 ) Transfer catalyzed by E1 Mechanism of redox

9. Step 3: transfer Maintenance of high energy bond Acetyl CoA product is made Lipoamide still reduced—not catalytically viable at this point

10. Dihydrolipoamide dehydrogenase (E 3 ) Redox of prosthetic FAD/FADH 2 Still not a regenerated catalyst!

11. Step 5: NADH produced Prosthetic group is restored Step 1 uses proton, step 5 regenerates Oxidation of one carbon atom used to – Produce high energy thioester – Produce NADH

12. Overall Reaction

13. Fate of Acetyl CoA Citric Acid Cycle (in muscle)—energy production (high potential electrons) Other tissues use intermediates in many ways – Amphibolic

14. Citric Acid Cycle Major points: carbon cycle, reaction types, reaction logic, energy harvest, selected enzyme reactions Minor points: enzyme names, substrate names, order of reactions

15. Energy Flow Decarboxylation or alcohol oxidation = NADH Double bond formation = QH 2 Dehydrogenases High energy bond = substrate phosphorylation

16. ATP Harvest: Net equations

17. Net ATP Harvest from Glucose Glycolysis = 2 ATP – Plus 3 or 5 ATP from NADH – In humans, cytosolic NADH leads to production of 3 ATP Pyruvate DH = 5 ATP Citric Acid Cycle = 20 ATP Total: 30 ATP/glucose in humans

18. Answers 1.A 2.C 3.B 4.D 5.A