2. Below is an image of the NADH cofactor, labeled A. (a) What role does this cofactor play, and draw a mechanism that connects the various states this molecule cycles between as part of its biological role. (b) In addition, four molecules resembling NADH are shown (labeled B-E). Which of these could fulfill the same role as NADH? Which of these would act as inhibitor? Explain your reasoning.
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- 4. (a) courses (such as this one) that NADH produced through glycoly- °H °H! sis in the cytosol can be used to generate ATP in the mitochon- dria. Yet, if 7-(14C)-NADH (in which the nicotinamide C-7 position is radioactively labeled; see diagram on the right) is added to rat hepatocytes, no radioactivity appears in the mitochondrial matrix while, if 4-(H)-NADH is added to hepatocytes, the tritium label soon appears in the mitochondrial matrix. Explain. It is generally stated and taught in biochemistry H H `NH2 NH2 R R [4-°H]NADH [7-14C]NADH1. Consider the enzyme pyruvate carboxylase. a. What pathway(s) does this enzyme function in? b. What cofactor does this enzyme use? c. Why is this cofactor required? d. How is this enzyme regulated? e. How does regulation of this enzyme impact the pathway(s) it operates in? How does regulation of this enzyme impact other pathway(s) we have discussed?2B. Calculate the total number of ATP that will be generated from the complete B-oxidation of oncobic acid (CH:(CH,),CH=CH(CH,),COOH; 15:1 cis-A9) in an organism that has all the standard required enzymes for metabolism but which pumps only 2 H" in complex I, 0 H' in complex II, 2 H* in complex III and 4 H' in complex IV. The F,F, ATPase is composed of 3 a/ß subunits and 10 c subunits. Show all of your calculations. HO
- 1. If the diagram below represents a catabolic pathway, and enzyme 1 requires NAD +, a) what type of reaction does enzyme 1 catalyze? b) what do coenzymes provide during enzymatic reactions? c) what are the roles of stoichiometric and catalytic coenzymes/cofactors? Enzyme 1 Enzyme 2 A B C W X Y z 2. If Z = ATP formed via substrate level phosphorylation in the diagram below, a) which of the following compounds (represented by an uppercase letter) contains a high-energy bond? b) compare and contrast substrate level phosphorylation and oxidative phosphorylation Enzyme 1 Enzyme 2 A W X Y Z11. Refer to the figure below. нн Н `NH2 NH2 N' N- 2e-+H* R NAD+ NADH NAD+ functions as a coenzyme in many enzyme-catalyzed reactions. The changes that take place in this coenzyme are the same for all of these reactions and are illustrated in the figure. It is likely that, in these reactions, NAD+ functions as an electron acceptor (reducing agent) in redox reactions. functions as an electron donor (oxidizing agent) in redox reactions. functions as a base in acid-base catalytic mechanisms. functions as an electron donor (oxidizing agent) in redox reactions. functions as an electron acceptor (oxidizing agent) in redox reactions. +Z-3. The nicotinamide coenzymes (see Fig. 13-24) can undergo reversible redox reactions with specific substrates in the presence of the appropriate redox enzyme. In these reactions NADH + H* serves as the hydrogen source as described in problem 1 above. In the reactions below, IF a redox change has occurred balance the reaction with the necessary amounts of NAD+, NADH, H* and H2O. The objective is to recognize when a redox enzyme is necessary in a metabolic reaction. (adapted from Problem 5, chapter 16) (a) CH,CH,OH CH, H. Ethanol Acetaldehyde (d) CH,- CH, + co2 OPO OH OPo? OH сH, — с —с `OPO;- Pyruvate Acetate (b) CH,-C-C + HPO? он TH. H. (e) -оос— сн, —с— соо- -оос— сн, — с — соо- 1,3-Bisphosphoglycerate Glyceraldehyde 3-phosphate H Oxaloacetate Malate (c) сH, — с —с сн, —с + co2 сH, — с — сн- + H* + CH,- C-CH; + Co, (f) Pyruvate Acetataldehyde Acetoacetate Acetone
- The following diagram shows reaction curves for aspartate transcarbamoylase (ATCase) with carbamoylphosphate and different concentrations of aspartate, in the absence of ATP (curve 1) and presence of ATP (curve 2). What do the shapes of the curves tell us about the ATCase enzyme? 2 جر [aspartate] It binds substrate through a sequential mechanism. It binds substte cooperatively. It obeys Michaelis-Menten kinetics. It binds substrate through a concerted mechanism.3. Answer the following questions about the metabolic pathway shown below: glutamate dehydrogenase e NH3 0-C-C-cH2-CH2-C- 0-C-CH-CH2-CH2-C-O + H,O + NAD + NH + NADH + H (a) Label the correct substances as the substrate, enzyme, and co-enzyme. (b) Which of the six classes does the enzyme of this reaction belong to? Why? (c) What is the name of the first molecule in this reaction? (d) Which metabolic pathway is this reaction likely to be a part of? A. glycolysis B. deamination C. beta-oxidation D. fermentation1. (a) The reaction catalyzed by citrate synthase is the first step of the TCA cycle. In glycolysis, two key reactions to produce ATP occur because an unfavorable reaction is coupled to another reaction that is thermodynamically favorable. The reaction catalyzed by citrate synthase is similarly coupled to an unfavorable reaction in the TCA cycle. Write the unfavorable TCA reaction using structural formulas and write the key step that drives the two coupled reactions forward. What is the overall AG" of the coupled reactions? (b). K(yM) 25.7 Inhibitor Bromoacetyl-CoA ATP NADH 6800 8300 The inhibitor constants for three inhibitors of por- cine citrate synthase are summarized in the table on the right. The compounds were all determined to bind in the ac- tive site as competitive inhibitors of acetyl-CoA. Because they bind as competitive inhibitors, all three inhibitors must exhibit structural similarity to some part of acetyl-CoA. Look up in the textbook the structural formulas for…
- 1. True / False: NADH dehydrogenase is the least electronegative complex of the ETC. 2. True / False: Most of the energy lost during the conversion of one form to another is as light. 3. True / False: The addition of a phosphate group is called phosphorylation. 4. True / False: The ETC pumps protons into the mitochondrial matrix. 5. True / False: FADH₂ transfers its electrons directly to cytochrome c. 6. True / False: FADH₂ is created exclusively during the citric acid cycle. 7. True/False: Alcohol fermentation occurs most commonly in animals and plants. 8. True / False: During fermentation, NAD* is regenerated when NADH is reduced. 9. True / False: The hydrolysis of ATP releases free energy. 10. True / False: Most of the CO₂ produced by cellular respiration is formed during ETC/chemiosmosis.10. Consider the beta oxidation of stearic acid (C18:0): How many ATP are generated in complete oxidation of stearic acid? How many NADH are generated in complete oxidation of stearic acid? How many FADH2 are generated in complete oxidation of stearic acid?1. The enzyme lactate dehydrogenase catalyzes the reaction below. NADH + H* NAD+ OH Pyruvate Lactate NADH absorbs light at 340 nm in the near-ultraviolet region of the electromagnetic spectrum, but NAD' does not. a. Assuming that you have a spectrophotometer that can measure light at this wavelength, qualitatively sketch a plot of A, 340 nm VS. time. How does this plot compare to the one from the experiment on the alkaline phosphatase-catalyzed reaction of p-nitrophenylphosphate? b. Would you use a highly accurate pH meter to monitor the progress of the pyruvate reaction? Why or why not?
