A) citrate to isocitrate. B) fumarate to malate. C) malate to oxaloacetate. D) succinate to fumarate. E) succinyl-coa to succinate

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1 Section 9 Key 1- There is reciprocal regulation of glycolytic and gluconeogenic reactions interconverting fructose-6-phosphate and fructose-1,6-bisphosphate. Which one of the following statements about this regulation is not correct? A) Fructose-2,6-bisphosphate activates phosphofructokinase-1. B) Fructose-2,6-bisphosphate inhibits fructose-1,6-bisphosphatase. C) The fructose-1,6-bisphosphatase reaction is exergonic. D) The phosphofructokinase-1 reaction is endergonic. E) This regulation allows control of the direction of net metabolite flow through the pathway. 2- The two moles of CO 2 produced in the first turn of the citric acid cycle have their origin in the: A) carboxyl and methylene carbons of oxaloacetate B) carboxyl group of acetate and a carboxyl group of oxaloacetate. C) carboxyl group of acetate and the keto group of oxaloacetate. D) two carbon atoms of acetate. E) two carboxyl groups derived from oxaloacetate. 3- The reaction of the citric acid cycle that produces an ATP equivalent (in the form of GTP) by substrate level phosphorylation is the conversion of: A) citrate to isocitrate. B) fumarate to malate. C) malate to oxaloacetate. D) succinate to fumarate. E) succinyl-coa to succinate 4- The standard reduction potentials (E' ) for the following half reactions are given. Fumarate + 2H + + 2e succinate FAD + 2H + + 2e FADH2 E' = V E' = V If succinate, fumarate, FAD, and FADH 2, all at l M concentrations, were mixed together in the presence of succinate dehydrogenase, which of the following would happen initially? A) Fumarate and succinate would become oxidized; FAD and FADH 2 would become reduced. B) Fumarate would become reduced; FADH 2 would become oxidized. C) No reaction would occur because all reactants and products are already at their standard concentrations. D) Succinate would become oxidized; FAD would become reduced. E) Succinate would become oxidized; FADH 2 would be unchanged because it is a cofactor, not a substrate.

2 5. Which of the following intermediates of the citric acid cycle is prochiral? A) Citrate B) Isocitrate C) Malate D) Oxaloacetate E) Succinate 6. Entry of acetyl-coa into the citric acid cycle is decreased when: A) [AMP] is high. B) NADH is rapidly oxidized through the respiratory chain. C) the ratio of [ATP]/[ADP is low D) the ratio of [ATP]/[ADP] is high. E) the ratio of [NAD + ]/[NADH] is high. 7. Citrate synthase and the NAD + -specific isocitrate dehydrogenase are two key regulatory enzymes of the citric acid cycle. These enzymes are inhibited by: A) acetyl-coa and fructose 6-phosphate. B) AMP and/or NAD +. C) AMP and/or NADH. D) ATP and/or NAD +. E) ATP and/or NADH. 8. If electron transfer in tightly coupled mitochondria is blocked (with antimycin A) between cytochrome b and cytochrome c1, then: A) all ATP synthesis will stop. B) ATP synthesis will continue, but the P/O ratio will drop to one. C) electron transfer from NADH will cease, but O 2 uptake will continue. D) electron transfer from succinate to O 2 will continue unabated. E) energy diverted from the cytochromes will be used to make ATP, and the P/O ratio will rise. 9. Which of the following statements about the chemiosmotic theory is correct? A) Electron transfer in mitochondria is accompanied by an asymmetric release of protons on one side of the inner mitochondrial membrane. B) It predicts that oxidative phosphorylation can occur even in the absence of an intact inner mitochondrial membrance. C) The effect of uncoupling reagents is a consequence of their ability to carry electrons through membranes. D) The membrane ATP synthase has no significant role in the chemiosmotic theory. E) All of the above are correct.

3 10. 2,4-Dinitrophenol and oligomycin inhibit mitochondrial oxidative phosphorylation. 2,4-Dinitrophenol is an uncoupling agent; oligomycin blocks the ATP synthesis reaction itself. Therefore, 2,4-dinitrophenol will: A) allow electron transfer in the presence of oligomycin. B) allow oxidative phosphorylation in the presence of oligomycin. C) block electron transfer in the presence of oligomycin. D) diminish O 2 consumption in the presence of oligomycin E) do none of the above. Short Answer: 11. Why is citrate, in addition to being a metabolic intermediate in aerobic oxidation of fuels, an important control molecule for a variety of enzymes? Ans: As the key biochemical intermediate in the citric acid cycle resulting from the condensation of oxaloacetate and acetyl-coa, citrate is at a junction of amino acid, fatty acid, and pyruvate oxidation, serving as an intracellular signal that the cell s current energy needs are being met. In particular, it is an allosteric regulator of PFK-1, increasing the inhibitory effect of ATP, and further reducing the flow of glucose through glycolysis. 12. Under what circumstances does the bifunctional protein phosphofructokinase-2/fructose 2,6-bisphosphatase (PFK-2/FBPase-2) become phosphorylated, and what are the consequences of its phosphorylation to the glycolytic and gluconeogenic pathways? Ans: Glucagon, signaling low blood sugar, stimulates camp synthesis, which activates protein kinase A (PKA) to phosphorylate PFK-2/FBPase-2 (among other proteins). This phosphorylation enhances FBPase-2 activity and inhibits PFK-2 activity of the enzyme, resulting in lower levels of fructose 2,6-bisphosphate (F26BP). In the absence of F26BP as an allosteric effector, the activity of PFK-1 is reduced (inhibiting glycolysis) and the activity of FBPase-1 is enhanced (stimulating gluconeogenesis), thus enabling the liver to replenish blood glucose. See Figs and The citric acid cycle begins with the condensation of acetyl-coa with oxaloacetate. Describe three possible sources for the acetyl-coa. Ans: Acetyl-CoA is produced by (1) the pyruvate dehydrogenase complex, (2) β oxidation of fatty acids, or (3) degradation of certain amino acids.

4 14. What is the function of FAD in the pyruvate dehydrogenase complex? How is it regenerated? Ans: FAD serves as the electron acceptor in the re-oxidation of the cofactor dihydrolipoate. It is converted to FADH 2 by this reaction and is regenerated by the passage of electrons to NAD During electron transfer through the mitochondrial respiratory chain, the overall reaction is: NADH + 1/2 O 2 + H + NAD + + H 2 O. The difference in reduction potentials for the two half-reactions ( E' ) is V. Show how you would calculate the standard free-energy change, G', for the reaction as written above. (The Faraday constant, I, is kj/v mol.) Ans: G' = ni E' = ( 2)(96.48 kj/v mol)(1.14v) = 220 kj/mol 16. When the F1 portion of the ATP synthetase complex is removed from the mitochondrial membrane and studied in solution, it functions as an ATPase. Why does it not function as an ATP synthetase? Ans: Like all enzymes, the F1 subunit of the ATP synthase catalyzes a reaction in both directions: ADP + Pi ATP + H 2 O The standard free-energy change ( G' ) for ATP hydrolysis is 30.5 kj/mol. With no proton motive force to drive the reaction toward ATP synthesis, the hydrolysis (ATPase activity) occurs spontaneously. 17. Using a simple diagram of the chemiosmotic theory, explain why anything that makes the mitochondrial membrane leaky stops ATP synthesis in the mitochondria. Ans: There are three central elements in the chemiosmotic model: (1) Electron flow through asymmetrically arranged membrane-bound carriers causes transmembrane flow of H +, creating a proton gradient (a proton motive force). (2) The proton motive force drives protons back across the membrane via specific proton channels (composed of Fo). (3) The energy released by downhill movement of protons is captured when ADP and Pi are condensed by ATP synthase (FoF1). Anything that makes the membrane leaky to protons (an uncoupler such as 2,4-dinitrophenol, or mechanical breakage of the membrane) prevents formation of a proton gradient. With no proton gradient, there is no energy source for ATP synthesis by FoF1 (ATP synthase). (See Fig , p. 705, and Fig , p. 717.)

5 18. Although molecular oxygen (O 2 ) does not participate directly in any of the reactions of the citric acid cycle, the cycle operates only when O 2 is present. Explain this observation. Ans: The citric acid cycle produces NADH, which is normally reoxidized to NAD+ by the passage of electrons through the respiratory chain to O 2. With no O 2 to accept electrons, NADH accumulates, NAD + is depleted, and the citric acid cycle slows for lack of NAD +.

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