Oxidative Phosphorylation Flashcards

(36 cards)

1
Q

What fuels ATP synthesis?

A

NADH and FADH2 bioenergy electron transfer to respiratory chain fuels ATP synthesis in mitochondria

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2
Q

What occurs in the Electron Transport Chain?

A

Oxidation of reduced coenzymes NADH and FADH2
Pumping of protons (H+)
Synthesis of ATP
Occurs in mitochondria

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3
Q

What is present in the diff parts of the mitochondria with regards to ETC?

A

Outer membrane - freely permeable to small molecules and ions

Inner membrane - impermeable to most small molecules and ions including H+
-Contains: respiratory electron carrieris (complexes I-IV), ADP-ATP translocase, ATP synthase (F0F1), other membrane transporters

Matrix - contains pyruvate dehydrogenase xomplex, citric acid cycle enzymes, fatty acid ß-oxidation enzymes, amino acid oxidation enzymes, DNA, ribosomes, many other enzymes, ATP, ADP, P, Mg2+, Ca2+, K+

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4
Q

Overview of what happens in oxidative phosphorylation?

A

Involves proteins and electron carriers that are part of the inner mitochondrial membrane
-Electrons from reduced coenzymes are removed and passed along a series of membrane-bound carriers
-The movement of electrons is a sequence of REDOX reactions
-The redox reactions are exergonic - the free energy made available by the reactions is used to punmp protons (H+) to the intermembrane space to establish a potential gradient
-Mkaing ATP rectifies the deficit in gradient by providing a channel through which the protons can pass back out into the matrix

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5
Q

How many protein complexes are involved in the ETC and where are they?

A

5 protein complexes
Located on inner mitochondrial membrane

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6
Q

How is ATP made by oxidative phosphorylation?

A

-Cells úse the energy from a flow of electrons through 4 protein complexes (respiratory chain) to pump H+ ions across a membrane
-The energy of the H+ gradient is harnessed to make ATP
-This is done in the presence of oxygen

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7
Q

What are I-IV enzyme complexes?

A

i NADH dehydrogenase
ii Succinate dehydrogenase
iii Ubiquinone cytochrome c oxidoreductase
iv Cytochrome oxidase

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8
Q

What is oxidative phosphorylation?

A

The process of ATP formation due to electron transfer from NADH/FADH2 to O2

-all catabolism in presence of O2 converges to oxidative phosphorylation
-in euks, occurs in inner mitochondrial membrane where respiratory electron carrier proteins and ATP synthase reside

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9
Q

What is the PMF?

A

Proton Motive Force
-energy of electron transfer efficiently conserved in proton gradient
-creates electrical potential - inside membrane negatuve and outside membrane positive

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10
Q

What is ATP synthesis driven by?

A

-PMF proton flow back to matrix through ATP-synthase complex - Complex V

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11
Q

What are the 3 types of electron transfer?

A

-Direct transfer as electrons
-Transfer as hydrogen atom
-Transfer as hydride ion

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12
Q

What is the term reducing equivalent used to describe?

A

A single elctron equivalent transferred in oxidation reduction reaction

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13
Q

What are the major electron carriers in the process?

A

-NAD, FAD, FMN
-Ubiquinone
-Iron-Sulfur proteins
-Cytochromes

(iron-sulphur and cytochromes are 2 diff forms of iron containing proteins)

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14
Q

Structure of Ubiquinone? How is it reduced/oxidised?

A

-Also called coenzyme Q or Q lipid
-Benzene type structure with long isoprenoid side chain
-Quinione ring structure can accept 1 or 2 electrons in hydrogen
-So is reduced from Q to QH to QH2
-Can be oxidised to donate these electrons

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15
Q

What are cytochromes?

A

-Proteins with iron containing heme prosthetic groups
-Different classes a, b, c
-4 N atoms coordinate to central iron
-The iron donates/accepts electrons; reduced/ oxidised; Fe2+/Fe
-a and b and some of c cclass part of integral membrane proteins but one form ogf cyt c is freely diffusible and soluble

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16
Q

What are Iron-sulphur proteins?

A

-iron complexed with sulphur
-S atoms in Cys residues of proteins - simple to complex structures
-As in cytochromes, the iron can be oxidised and reduced - one elctron transfer
-Atleast 8 of these work in the ETC

17
Q

Electron carrier sequence?

A

NADH
Flavoprotein
Ubiquinone
Fe-S centres
Cytochromes
Finally O2

18
Q

How is the sequence of the electron carriers determined?

A

By use of inhibitors that can block electron transport between the carriers

19
Q

What are electrons carried by to complete oxidative phosphorylation?

A

Electrons are carried via electron carriers in supramolecular enzyme complexes embedded in inner mitochondrial membrane
-Complex I-IV - together known as respiratory chain
-Also contains mobile electron carriers unbiquinone and Cytochrome C

20
Q

Where is the respiratory chain found?

A

Inner membrane of mitochondria - within cristae

21
Q

What is and what does Complex I do?

A

NADH dehydrogenase
-very large with 45 diff polypeptide chains
-transfers 2 electrons from NADH to ubiquinone (Q)
NADH -> NAD+ + Q -> QH2
-4 protons H+ pumped across matrix –> intermembrane space

22
Q

What is and what does Complex II do?

A

Succinate Dehydrogenase
-also part of citric acid cycle
-transfers electrons from succinate -> FADH2 -> ubiquinone
so electrons pass from succinate ->FAD -> FADH2 -> FAD+ + Q-> QH2

23
Q

What is and what does Complex III do?

A

Ubiquinone Cytochrome C oxidoreductase performs very complex electron transfer= Q cycle
1. end result is UQH2 transfers 2 electrons to two Cytochrome C
2. Electron trasnfer drives expulsion of 4 more prtons from matrix to intermembrane space

24
Q

What is and what does Complex IV do?

A

Cytochrome Oxidase
-Transfers electrons from cytochrome C to O2 - complete electron transfer
-Electrons trnsferred from Cyt C -> Cyt a -> Cyt a3 -> O2
1. carries out 4 electron reduction of O2 to H2O
2. 2 protons transferred from matrix to intermembrane space

25
Inhibitors that block electron transport from electron carriers?
-Rotenonne blocks b/w Complex I and Q -Antimycin blocks b/w Complex III and IV -Cyanide blocks b/w Complex IV and O2
26
What does each NADH that donates electrons to respiratory chain drive the expulsion of?
4H+ through complex I and III and 2H+ through complex IV -10 H+ travel to intermembrane space
27
What does each FADH2 that donates electrons to respiratory chain drive the expulsion of?
4H+ through complex III and 2H+ through complex IV -6 H+ travel to intermembrane space (Electron transfer creates PMF - inside membrane matrix is negative and outside membrane matrix is positive)
28
What are other sources of FADH2
-FADH2 is not just produced by the succinate dehydrogenase rxn succinate to fumarate -Also produced from 1st rxn in fatty acid oxidation catalysed by Acyl-CoA dehydrogenase -Donates electrons from FADH2 to ETF (electron transfer flavoprotein) which donates electrons to ubiquinone
29
What did Peter Mitchell propose?
The chemiosmotic model -PMF conserves energy of electron transfer and drives ATP synthesis as protons flow back into mitochondrial matrix via a proton pore F0 in inner mitochondrial membrane -F0 is part of ATP synthase enzyme
30
What did his experiments show?
-Electron transfer, PMF and ATP synthesis are obligately coupled -Inhibitors of electron transport block ATP synthesis -Inhibitors of ATP synthesis block electron transport
31
What did John Walker get a Nobel prize for in 1977?
-Working out how PMF causes enzyme ATP synthase to synthesise ATP -'the ATP synthase complexes in mitochondria make the ATP required to sustain life by rotary mechanism' -ATP synthase enzyme = F-type ATPase purified and crystallised has 2 distinct functional domains F1 and F0 -human ATP synthase aasmbled from 27 nuclear encoded genes and 2 mitochondrially encoded genes
32
What is F1?
-Peripheral membrane protein -Attached to inner mitochondrial membrane -N side -Catalytic domain that synthesizes ATP
33
What is F0?
-integral membrane protein -spans mitochondrial inner membrane -proton pore -H+ move through it againts conc gradient from intermembrane space through F1 to matrix -Proton flow enables ATP synthesis
34
F1 structure?
-9 subunits - 5 diff types -composition - alpha 3 ß 3 y delta e -alternating alpha and ßs surround central y shaft -alpha and beta subunits have nucleotide binding sites -ß subunits more important role can bind ADP, ATP or be empty
35
F0 structure?
-Integral membrane protein -makes up proton pore -composed of 3 subunits a b2 cn -C subunits cyclinder-like structure with pore - number of C subunits differs b/w species 8-15 -C subunits rotate together around perpendicular axis - around e and y which `stand`on C subunits
36
What did Paul Boyer do?
-Proposed the binding change mechanism to explain how ATP synthase synthesises ATP -protons pass through F0 and cause cylinder of c units attached to y - subunit of F1 to rotate about long axis of y -y-subunit rotates and passes through centre of alpha3 ß3 which are stationary -rotational catalysis causes ATP synthesis