2. Mitochondria isolated from bovine cardiac muscle, when subjected to sonication, form closed mem- brane vesicles containing the electron transport catalysts and the ATP synthase enzyme located in the inner mitochondrial membrane. A diagrammatic representation of these electron transport particles retaining the capacity for oxidative phosphorylation is given below. NADH- NAD+ tion? H₂O ATP 1/202 NADH Q₁H₂ NAD+ Complex I Complex III ADP Complex IV ADP ATP In the diagram protein complexes colored gray indicate that they are inhibited while complexes colored red, green, etc. are catalytically active. Complex II (a) ( In Panel A indicate which side of the membrane vesicle faced the matrix and which side faced the intermembrane space prior to sonication. During coupled substrate oxidation which compart- ment will be of higher pH and which of lower pH. (b) For the system in Panel A, sonication of mitochondria was carried out in the presence of an excess of ferricytochrome c (i.e., the heme iron is in the ferric state) and Coenzyme Q (CoQ or ubiquinone) enabling both to be trapped into the vesicular cavity upon closing of membrane fragments into spherical vesicles. For oxidation of NADH by O2 coupled to phosphorylation of ADP, in addition to NADH and ADP, what other reagent must be added to the suspension of electron transport particles to observe oxidative phosphorylation and how does this differ from the corresponding process in intact mitochondria?. Why would addition of oligomycin not alter the coupling of oxidation and phosphoryla-

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
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Chapter1: Biochemistry: An Evolving Science
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2. Mitochondria isolated from bovine cardiac muscle, when subjected to sonication, form closed mem-
brane vesicles containing the electron transport catalysts and the ATP synthase enzyme located in the
inner mitochondrial membrane. A diagrammatic representation of these electron transport particles
retaining the capacity for oxidative phosphorylation is given below.
NADH
NAD+
(
ATP
tion?
H₂O
ADP
1/202
NADH
Q₁H₂
NAD+
Q₁
Complex I
Complex III
ADP
Complex IV
ATP
In the diagram protein complexes colored gray indicate that they are inhibited while complexes colored
red, green, etc. are catalytically active.
Complex II
(a)(
In Panel A indicate which side of the membrane vesicle faced the matrix and which side
faced the intermembrane space prior to sonication. During coupled substrate oxidation which compart-
ment will be of higher pH and which of lower pH.
(b)
For the system in Panel A, sonication of mitochondria was carried out in the presence of
an excess of ferricytochrome c (i.e., the heme iron is in the ferric state) and Coenzyme Q (CoQ or
ubiquinone) enabling both to be trapped into the vesicular cavity upon closing of membrane fragments
into spherical vesicles. For oxidation of NADH by O2 coupled to phosphorylation of ADP, in addition to
NADH and ADP, what other reagent must be added to the suspension of electron transport particles to
observe oxidative phosphorylation and how does this differ from the corresponding process in intact
mitochondria?. Why would addition of oligomycin not alter the coupling of oxidation and phosphoryla-
Transcribed Image Text:2. Mitochondria isolated from bovine cardiac muscle, when subjected to sonication, form closed mem- brane vesicles containing the electron transport catalysts and the ATP synthase enzyme located in the inner mitochondrial membrane. A diagrammatic representation of these electron transport particles retaining the capacity for oxidative phosphorylation is given below. NADH NAD+ ( ATP tion? H₂O ADP 1/202 NADH Q₁H₂ NAD+ Q₁ Complex I Complex III ADP Complex IV ATP In the diagram protein complexes colored gray indicate that they are inhibited while complexes colored red, green, etc. are catalytically active. Complex II (a)( In Panel A indicate which side of the membrane vesicle faced the matrix and which side faced the intermembrane space prior to sonication. During coupled substrate oxidation which compart- ment will be of higher pH and which of lower pH. (b) For the system in Panel A, sonication of mitochondria was carried out in the presence of an excess of ferricytochrome c (i.e., the heme iron is in the ferric state) and Coenzyme Q (CoQ or ubiquinone) enabling both to be trapped into the vesicular cavity upon closing of membrane fragments into spherical vesicles. For oxidation of NADH by O2 coupled to phosphorylation of ADP, in addition to NADH and ADP, what other reagent must be added to the suspension of electron transport particles to observe oxidative phosphorylation and how does this differ from the corresponding process in intact mitochondria?. Why would addition of oligomycin not alter the coupling of oxidation and phosphoryla-
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