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4. The diagram below illustrates time dependent O₂ uptake by isolated skeletal muscle mito- chondria under various conditions that affect the rate of respiration. The following abbreviations with arrows indicate when addition of various ingredients is made to the reaction chamber: The vertical, red lines have been added to help delineate time intervals corresponding to addition and metabolic con- version of ADP. Abbreviations: Med addition of 0.3 M sucrose; Pi: addition of 0.01 M phosphate buffer, pH 7.4; Mw: addition of isolated, intact mitochondria; Pyr + Mal: addition of 0.01 M pyruvate + 0.01 M malate; ADP : addition of adenosine 5'-diphosphate; Oligo/mg: addition of oligomycin/mg protein; FCCP : addition of the uncoupler FCCP. Med Pyr+Mal P₁ 190μM ADP ↓ 20μM 02 T H 190μM ADP I Minute 475μM ADP 2μM FCCP 13μg Oligo/mg -[0₂] = 0 ATP [ATP] = 0 ApH = (pH matrix -pHout) ApH = 0 Start Finish (a) Why is there an increase in the rate of O2 uptake for the first two additions of ADP and why does the rate return to the resting level? (b) Why is a combination of pyruvate and malate added to the mitochondria to support respira- tion rather than pyruvate alone? (c). ) In the row marked ATP in the diagram above, plot the total, relative amount of ATP generated in the mitochondria as a function of time from start to finish, indicating how it changes based on the sequential addition of metabolites, ADP, oligomycin, and the uncoupler FCCP. (d) Similarly, in the row marked ApH = (pHmatrix-pHout) indicate when build-up or annihilation of the proton gradient occurs as an all-or-none phenomenon using the same time scale for the addi- tion B4-

4. The diagram below illustrates time dependent O₂ uptake by isolated skeletal muscle mito- chondria under various conditions that affect the rate of respiration. The following abbreviations with arrows indicate when addition of various ingredients is made to the reaction chamber: The vertical, red lines have been added to help delineate time intervals corresponding to addition and metabolic con- version of ADP. Abbreviations: Med addition of 0.3 M sucrose; Pi: addition of 0.01 M phosphate buffer, pH 7.4; Mw: addition of isolated, intact mitochondria; Pyr + Mal: addition of 0.01 M pyruvate + 0.01 M malate; ADP : addition of adenosine 5'-diphosphate; Oligo/mg: addition of oligomycin/mg protein; FCCP : addition of the uncoupler FCCP. Med Pyr+Mal P₁ 190μM ADP ↓ 20μM 02 T H 190μM ADP I Minute 475μM ADP 2μM FCCP 13μg Oligo/mg -[0₂] = 0 ATP [ATP] = 0 ApH = (pH matrix -pHout) ApH = 0 Start Finish (a) Why is there an increase in the rate of O2 uptake for the first two additions of ADP and why does the rate return to the resting level? (b) Why is a combination of pyruvate and malate added to the mitochondria to support respira- tion rather than pyruvate alone? (c). ) In the row marked ATP in the diagram above, plot the total, relative amount of ATP generated in the mitochondria as a function of time from start to finish, indicating how it changes based on the sequential addition of metabolites, ADP, oligomycin, and the uncoupler FCCP. (d) Similarly, in the row marked ApH = (pHmatrix-pHout) indicate when build-up or annihilation of the proton gradient occurs as an all-or-none phenomenon using the same time scale for the addi- tion B4-

Biochemistry
Biochemistry
6th Edition
ISBN:9781305577206
Author:Reginald H. Garrett, Charles M. Grisham
Publisher:Reginald H. Garrett, Charles M. Grisham
Chapter21: Photosynthesis
Section: Chapter Questions
Problem 5P: The Relative Efficiency of ATP Synthesis in Noncyclic versus Cyclic Photophosphorylation If...
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4. The diagram below illustrates time dependent O₂ uptake by isolated skeletal muscle mito- chondria under various conditions that affect the rate of respiration. The following abbreviations with arrows indicate when addition of various ingredients is made to the reaction chamber: The vertical, red lines have been added to help delineate time intervals corresponding to addition and metabolic con- version of ADP. Abbreviations: Med addition of 0.3 M sucrose; Pi: addition of 0.01 M phosphate buffer, pH 7.4; Mw: addition of isolated, intact mitochondria; Pyr + Mal: addition of 0.01 M pyruvate + 0.01 M malate; ADP : addition of adenosine 5'-diphosphate; Oligo/mg: addition of oligomycin/mg protein; FCCP : addition of the uncoupler FCCP. Med Pyr+Mal P₁ 190μM ADP ↓ 20μM 02 T H 190μM ADP I Minute 475μM ADP 2μM FCCP 13μg Oligo/mg -[0₂] = 0 ATP [ATP] = 0 ApH = (pH matrix -pHout) ApH = 0 Start Finish (a) Why is there an increase in the rate of O2 uptake for the first two additions of ADP and why does the rate return to the resting level? (b) Why is a combination of pyruvate and malate added to the mitochondria to support respira- tion rather than pyruvate alone? (c). ) In the row marked ATP in the diagram above, plot the total, relative amount of ATP generated in the mitochondria as a function of time from start to finish, indicating how it changes based on the sequential addition of metabolites, ADP, oligomycin, and the uncoupler FCCP. (d) Similarly, in the row marked ApH = (pHmatrix-pHout) indicate when build-up or annihilation of the proton gradient occurs as an all-or-none phenomenon using the same time scale for the addi- tion B4-
Transcribed Image Text:4. The diagram below illustrates time dependent O₂ uptake by isolated skeletal muscle mito- chondria under various conditions that affect the rate of respiration. The following abbreviations with arrows indicate when addition of various ingredients is made to the reaction chamber: The vertical, red lines have been added to help delineate time intervals corresponding to addition and metabolic con- version of ADP. Abbreviations: Med addition of 0.3 M sucrose; Pi: addition of 0.01 M phosphate buffer, pH 7.4; Mw: addition of isolated, intact mitochondria; Pyr + Mal: addition of 0.01 M pyruvate + 0.01 M malate; ADP : addition of adenosine 5'-diphosphate; Oligo/mg: addition of oligomycin/mg protein; FCCP : addition of the uncoupler FCCP. Med Pyr+Mal P₁ 190μM ADP ↓ 20μM 02 T H 190μM ADP I Minute 475μM ADP 2μM FCCP 13μg Oligo/mg -[0₂] = 0 ATP [ATP] = 0 ApH = (pH matrix -pHout) ApH = 0 Start Finish (a) Why is there an increase in the rate of O2 uptake for the first two additions of ADP and why does the rate return to the resting level? (b) Why is a combination of pyruvate and malate added to the mitochondria to support respira- tion rather than pyruvate alone? (c). ) In the row marked ATP in the diagram above, plot the total, relative amount of ATP generated in the mitochondria as a function of time from start to finish, indicating how it changes based on the sequential addition of metabolites, ADP, oligomycin, and the uncoupler FCCP. (d) Similarly, in the row marked ApH = (pHmatrix-pHout) indicate when build-up or annihilation of the proton gradient occurs as an all-or-none phenomenon using the same time scale for the addi- tion B4-
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