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- (c) (1 ) Fluoroacetate, prepared commercially for rodent control, is also produced by a South Afri- can plant. After entering a cell, fluoroacetate is converted to fluoroacetyl-CoA in a reaction catalyzed by the enzyme acetate thiokinase: F-CH2COO¯ + COA-SH + ATP F-CH2CO-SCOA + AMP + PPj The toxic effect of fluoroacetate was studied in an experiment using intact isolated rat heart. After the heart was perfused with 0.22 mM fluoroacetate, the measured rate of glucose uptake and glycolysis decreased, and glucose 6-phosphate and fructose 6-phosphate accumulated. Examination of the cit- ric acid cycle intermediates revealed that their concentrations were below normal, except for citrate, with a concentration 10-fold higher than normal. Where did the block in the citric acid cycle occur? What caused citrate to accumulate and the other cycle intermediates to be depleted? Explain. Why does it block the citric acid cycle? Fluoroacetyl-CoA is enzymatically transformed in the citric acid cycle.…The formation of acetyl-CoA from acetate is an ATP-driven reaction: Acetate + ATP + CoA Acetyl CoA + AMP + PP₁ Calculate AG" for this reaction given that the AG' for the hydrolysis of acetyl CoA to acetate and CoA is −31.4 kJ mol-¹ (−7.5 kcal mol¯¹) and that the AG°' for hydrolysis of ATP to AMP and PP; is -45.6 kJ mol-¹ (−10.9 kcal mol−¹ ). AG° reaction = AGO! i The PP, formed in the preceding reaction is rapidly hydrolyzed in vivo because of the ubiquity of inorganic pyrophosphatase. The AG" for the hydrolysis of pyrophosphate (PP;) is −19.2 KJ mol-¹ (-4.665 kcal mol-¹). Calculate the AGº' for the overall reaction, including pyrophosphate hydrolysis. reaction with PP, hydrolysis = What effect does the presence of pyrophosphatase have on the formation of acetyl CoA? kJ mol-¹ It does not affect the overall reaction. It brings the overall reaction closer to equilibrium. It makes the overall reaction even more exergonic. It makes the overall reaction even more endergonic. kJ mol-¹ -1(b) ☐ The exergonic reactions catalyzed by isocitrate dehydrogenase (ICDH) and α- ketoglutarate dehydroge-nase (aKGDH) are regu- lated by metabolites, in addition to the reaction catalyzed by citrate synthase. The diagram to the right illustrates the activity of ICDH (A,▲) and aKGDH (O,●) in the presence of 1.5 mM ADP (O,A) or 1.5 mM ATP (●,▲). In muscle tissue contraction of sarcomeres initiated by a nerve im- pulse results first in the release of Ca 2+ from the sarcoplasmic reticulum. As indicated by the chang- es in velocity of the two enzymes, what are the flux changes of the TCA cycle in muscle tissue upon release of Ca2+ under the influence of ADP viz. ATP and under the influence of NAD+ viz. NADH on cit- rate synthase? 100 75 Stimulation of activity (% of maximum) ŏ 25 1. -8 -7 -6 log {[Ca2+] (M)}
- The synthesis of the activated form of acetate (acetyl-CoA) is carried out in an ATP-dependent process: Acetate + CoA + ATP → acetyl-CoA + AMP + PPi (a) The ∆G’o for the hydrolysis of acetyl-CoA to acetate and CoA is - 32.2 kJ/mol and that for hydrolysis of ATP to AMP and PPi is - 30.5 kJ/mol. Calculate ∆G’o for the ATP-dependent synthesis of acetyl-CoA. (b) Almost all cells contain the enzyme inorganic pyrophosphatase, which catalyzes the hydrolysis of PPi to Pi. What effect does the presence of this enzyme have on the synthesis of acetyl-CoA? Explain.Citrate synthase catalyzes the reaction Oxaloacetate + acetyl-CoA →citrate + HS-CoA The standard free energy change for the reaction is −31.5 kJ · mol−1. (a) Calculate the equilibrium constant for this reaction at 37°C. (b) Would you expect this reaction to serve as a control point for its pathway (the citric acid cycle)?Fatty acids are converted to their coenzyme A esters in a reversible reaction catalyzed by acyl-CoA synthetase: R-COO +ATP +COA R-C-COA +AMP + PPi a) The reaction involves two steps the first of which forms an enzyme-bound intermediate identified as the mixed anhydride of the fatty acid and AMP: R-C-O-P-O-nibose-adenine Write two chemical equations coresponding to the two steps of the reaction catalyzed by the synthetase. b) The acyl-CoA synthetase reaction as written above is readily reversible. How might the reaction be made to favor formation of fatty acyl-CoA? Write within the box. Anything outside the box will not be graded. From the
- (i) Consider a preparation that contains all the enzymes and cofactors necessary for fatty acid biosynthesis from acetyl-CoA and malonyl-CoA. If [2-H] acetyl-CoA labeled with deuterium, the heavy isotope of hydrogen and excess of unlabeled malonyl-CoA are added as substrates, where will you find these labeled deuterium atoms in a molecule of palmitate synthesized? Explain. S-COA (ii) Describe the steps involved in the synthesis of palmitic acid starting from acetyl-CoA and malonyl-CoA.Given the following statements, identify which are true and which are not. (1) Two ribose subunits are present in the coenzyme flavin adenine dinucleotide. (2) In the common metabolic pathway, 10 molecules of ATP are produced per acetyl CoA catabolized. (3) The purpose of step 2 in the CAC is to convert a secondary alcohol to a tertiary alcohol.Synthesis of the activated form of acetate (acetyl-CoA) is carried out in an ATP-dependent process:(a) The ΔG′° for hydrolysis of acetyl-CoA to acetate and CoA is −32.2 kJ/mol and that for hydrolysis of ATP to AMP and PPi is −30.5 kJ/mol. Calculate ΔG′° for the ATPdependent synthesis of acetyl-CoA.(b) Almost all cells contain the enzyme inorganic pyrophosphatase, which catalyzes the hydrolysis of PPi to Pi. What effect does the presence of this enzyme have on the synthesis of acetyl-CoA? Explain.
- The complete oxidation of palmitoyl-CoA to carbon dioxide and water is presented by the overall equation: Palmitoyl-CoA + 23O2 + 108Pi + 108 ATP + 23H2O → CoA + 16 CO2 + 108 ATP + 23H2O Water is also produced in the reaction ADP + Pi → ATP + H2O But not included as a product in the overall equation. Why?9. The equation below depicts the first step of the citric acid cycle. H0 CoA-SH CH-C + 0=C-CO0 HO C-COO citrate s-COA ČH2-COO synthase ČH2 -CO0 Citrate Acetyl-CoA Oxaloacetate AG" = -32.2 kJ/mol a) Explain the chemical conversions that take place during this step. b) Why is this reaction energetically favorable? Explain.Page of 6 ZOOM + name: 3. In the last reaction of the citric acid cycle, malate is dehydrogenated to regenerate the oxaloacetate necessary for the entry of acetyl-CoA into the cycle: L-Malate + NAD+ → oxaloacetate + NADH + H* AG'° = 30.0 kJ/mol (a) Calculate the equilibrium constant for this reaction at 25 °C. (b) Because AG°' assumes a standard pH of 7, the equilibrium constant calculated in (a) corresponds to [oxaloacetate][NADH] Keq [L-malate][NAD*] The measured concentration of L-malate in rat liver mitochondria is about 0.20 mM when [NAD*]/[NADH] is 10. Calculate the concentration of oxaloacetate at pH 7 in these mitochondria. (c) To appreciate the magnitude of the mitochondrial oxaloacetate concentration, calculate the number of oxaloacetate molecules in a single rat liver mitochondrion. Assume the mitochondrion is a sphere of diameter 2.0 microns.