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Classic papers in biochemistry

Structure at 2.8 resolution of F1-ATPase from bovine heart mitochondria


Nature, 370, 1994, 621-628 Abrahams, Leslie, Lutter & Walker

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Mitochondria and F-ATPase

1980-1994

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How is pmf coupled to ATP synthesis?


Direct (Mitchell, Williams) Indirect (Boyer, others)

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The thirteen proteins encoded in mitochondrial DNA

Human mtDNA
7 proteins 1 protein 2 proteins 3 proteins

2NAD+ 2NADH
2 e

4H
+

FADH
2

FA D 2 e Q QH
2

2H
+

4H
+

H2 O

[O ]
2 e

ATP ADP + Pi

3-4 H+

Q QH
2

4H
+
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2 H+ e

2 e

3-4 H+ 2H
+

Cyt c

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P-loop sequences in F1-ATPase

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P-loop sequences in F1-ATPase


consensus 169-1 159-163 G X X X X G KT G D R QT G KT G G A GV G KT

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Paul Boyer

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ATP binding site of TP

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The ATP synthase

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20 nm

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Generation of rotation

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The H+/ATP and Na+/ATP ratios

For the synthesis of each ATP, the -subunit rotates through 120 12 c-subunits/ring



H+/ATP = 4 H+/ATP = 3.3 H+/ATP = 4.7

10 c-subunits in S. cerevisiae

14 c-subunits in chloroplasts

11 c-subunits in Na+-motive F-ATPases


Na+/ATP = 3.7 Estimates of H+/ATP are in the range 3-4

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The mammalian c-ring


The bioenergetic cost of making an ATP molecule in mammalian mitochondria

Watt et al (2010) PNAS 107, 16823

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P/O ratio in mammalian mitochondria NADH 2e- to O2 =10 H+ pumped out + Succinate 2e to O2 = 6 H pumped out ADP/ATP exchange + Pi/H+ symport = 1 H+ in ATP synthase ATP synthesis H /ATP = 8/3 = 2.7 H+/ATP = 3.7
+

Moles ADP phosphorylated/2e to O2 = P/O ratio e donor NADH Succinate


-

P/O calc 10/3.7=2.7 6/3.7=1.6

P/O exp 2.5 1.5

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Species known to have trimethyl-lysine-43

Man, cows, sheep, pigs, rabbit, mice, rats Chickens Flies (Diptera) NB Yeast has K43, but not trimethylated

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17

Lindsay Baker, Mike Runswick, John Walker and John Rubinstein, unpublished work
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Lindsay Baker, Mike Runswick, John Walker and John Rubinstein, unpublished work
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Unresolved issues

The a-c-ring interface Understanding differences between F-ATPases (regulation, ring symmetries, peripheral stalks) Reconciling structures and biophysics

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