25. The following graph illustrates the consumption of oxygen in a mitochondrial preparation. Draw, on the graph, what would happen to oxygen concentration in the mitochondria under each of the following conditions (in other words, your final graph should have 4 different lines on it, clearly labeled a, b, c, and d) A .3 dte: 0cT ON 01E.0 .2t .1 1 2 3 Time, min (a) NADH (but nothing else) is added at the time marked A (b) NADH plus ADP is added at the time marked A. (c) NADH plus ADP plus a drug that prevents ATP synthase from binding to ADP at the time marked A. (d) NADH plus ADP plus a drug that prevents ATP synthase from binding ADP at the time marked A, followed by a drug that makes the inner membrane space more basic 1 minute later. [02], mM

25. The following graph illustrates the consumption of oxygen in a mitochondrial preparation. Draw, on the graph, what would happen to oxygen concentration in the mitochondria under each of the following conditions (in other words, your final graph should have 4 different lines on it, clearly labeled a, b, c, and d) A .3 dte: 0cT ON 01E.0 .2t .1 1 2 3 Time, min (a) NADH (but nothing else) is added at the time marked A (b) NADH plus ADP is added at the time marked A. (c) NADH plus ADP plus a drug that prevents ATP synthase from binding to ADP at the time marked A. (d) NADH plus ADP plus a drug that prevents ATP synthase from binding ADP at the time marked A, followed by a drug that makes the inner membrane space more basic 1 minute later. [02], mM

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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1. Name and draw diagrammatically the series of mitochondrial electron transfer catalysts, starting with the oxidation of NADH and succinate and ending with the reduction of O2. 2. Indicate the sites and stoichiometry (per 2e) at which protons are translocated from the matrix to the intermembrane space. 3. Indicate which complexes are inhibited by: amytal, antimycin A, azide (N3 ), cyanide (CN), carbon monoxide (CO), and rotenone. 4. Describe the effects of (1) oligomycin and (2) uncouplers of oxidative phosphorylation, e.g., dinitrophenol (DNP), carbonyl cyanide-p- trifluoromethoxyphenylhydrazone (FCCP), on respiration and ATP synthesis when added to a suspension of mitochondria with excess malate, ADP, and inorganic phosphate (Pi).
Chambers and coworkers have reported [NAD+] and [NADH] concentrations in yeast mitochondria as 20 mM and 0.3 mM, respectively. Consider the Malate Dehydrogenase reaction below: Malate + NAD+ → Oxaloacetate + NADH + H+ ∆G0’ = +29.7 kJ/mol If Malate concentration in yeast mitochondria is 0.4 mM what is the maximum concentration of oxaloacetate needed to make the reaction exergonic at pH 7.0 and 370C?
11. Chambers and coworkers have reported [NAD*] and [NADH] concentrations in yeast mitochondria as 20 mM and 0.3 mM, respectively. Consider the Malate Dehydrogenase reaction below: Malate + NAD* → Oxaloacetate + NADH + H* AG° = +29.7 kJ/mol If Malate concentration in yeast mitochondria is 0.4 mM what is the maximum concentration of oxaloacetate needed to make the reaction exergonic at pH 7.0 and 37°C? А. 7.14 х 105 В. 2.04 х 104 С. 8.17 х 106 D. 5.45 x 108 Е. 2.63 х 107 F. 4.69 x 108 G. 4.90 X107 Н. 3.13 х 106 I. 5.05 х 106
PFor each of the following experiments, tell me whether you would expect the rate of O₂ consumption and ATP synthesis to increase, decrease or remain the same (respectively). Assume there is plenty of O₂ around. (a) Succinate is suddenly added to mitochondria that have no other oxidizable substrates (Assume that there are ADP and Pi in mitochondria). (b) ADP is not allowed to pass into the mitochondrial matrix. (c) 2,4 Dinitrophenol (an uncoupler) is added.
1. Cyanide, oligomycin, and 2,4-dinitrophenol are all inhibitors of oxidative phosphorylation in mitochon- dria. Provide an explanation to the following conditions regarding these potent inhibitors. (a) Explain why adding cyanide to an active in vitro suspension of mitochondria blocks ATP synthesis. What happens to the rate of ATP synthesis when 2,4-dinitrophenol is added to this mitochon- drial suspension after it was treated with cyanide? (b) Explain why the rate of oxygen consumption decreases in an in vitro suspension of mito- chondria when oligomycin is added. What happens to the rate of oxygen consumption in this oligomycin- inhibited system after adding 2,4-dinitrophenol? Explain.
7) The following questions apply to glycolysis (figure 3-41 on page 79 of the 12th edition), the Krebs cycle (figure 3-44 on page 82) and to oxidative phosphorylation (figure 3-45 on page 84). Calculate the total amount of ATP produced from one glucose molecule if: a. The oxidation of NADH + H* to NAD* + 2H* produced 4 ATP. b. The oxidation of FADH₂ to FAD + 2H* produced 1 ATP. c. Each step of the Krebs cycle that produces NADH+H* produced FADH2 instead. d. Each step of the Krebs cycle that produces FADH2 produced NADH + H+ instead. e. Sucrose is substituted for glucose.
(A) What is the free energy yield associated with the transport of a proton from the mitochondrial intermembrane space (IMS) to the mitochondrial matrix, assuming that the pH of the IMS is 1.4 units lower than the pH of the matrix? (Hint: Calculate AGO' for the reaction H"IMS > H matris using the relationship AG° = RTInKeg and remembering that pH = -log1o[H*]. Assume that T = 310K (37°C), and R = 8.315x10-3 kJ mol-1 K-1. Show your work and include correct units!)
You are isolating mitochondria from insect cells and incubating in a test tube with 0.005 M FADH2, 0.05 M ADP and 0.05 M Pi. Assuming, these 3 can enter mitochondria at no cost,no glucose/products of glucose metabolism remain in the isolated mitochondria, and oxygen is present. Part 1) If all expected reactions go to completion, how much ATP is expected formed? (0 M)(0.02 M) (0.03 M) (0.05 M) (0.1 M) (0.005 M) (0.01 M) (0.015 M) Part 2) The ratio of FADH2/FAD at completion of all expected reactions would be? (2) (>>2) (0) (1) Part 3) After completion of all expected reactions, ratio of H ion concentration inside vs. outside the mitochondrial inner membrane should be? (<1) (~1) (>1)(0) Please provide brief explanation
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]NADH
Make a rough drawing of the ATP synthase machinery within the mitochondrial membrane and specifically label the Fo and F1 subunits, as well as the C-ring, the a subunit, the y stalk and the a3ß3 hexamer.
When pure reduced cytochrome c is added to carefully prepared mitochondria along with ADP, P, antimycin A, and oxygen, the cytochrome c becomes oxidized, and ATP is formed, with a P/o ratio approaching 1.0. (a) Indicate the probable flow of electrons in this system. (b) Why was antimycin A added? (c) What does this experiment tell you about the location of cou- pling sites for oxidative phosphorylation? (d) Write a balanced equation for the overall reaction (including cyt c oxidation and ATP synthesis).
1 a) What is meant by the ATP currency exchange ratio? Why does the oxidation of mitochondrial FADH2 generate one less ATP than oxidation of mitochondrial NADH? b) If 12 H+ are moved across the inner mitochondrial membrane by NADH oxidation, and each ATP synthesized requires 3 H+ to move through ATP synthase, why are only 3 ATP molecules produced by oxidation of each NADH?