Large conformational changes of the ε subunit in the bacterial F1F0 ATP synthase provide a ratchet action to regulate this rotary motor enzyme

被引:157
作者
Tsunoda, SP
Rodgers, AJW
Aggeler, R
Wilce, MCJ
Yoshida, M
Capaldi, RA [1 ]
机构
[1] Univ Oregon, Inst Mol Biol, Eugene, OR 97403 USA
[2] Tokyo Inst Technol, Chem Res Lab R1, Yokohama, Kanagawa 2268503, Japan
[3] Univ Western Australia, Dept Pharmacol, Nedlands, WA 6907, Australia
[4] Univ Western Australia, Crystallog Ctr, Nedlands, WA 6907, Australia
关键词
D O I
10.1073/pnas.111128098
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The F1F0 ATP synthase is the smallest motor enzyme known. Previous studies had established that the central stalk, made of the gamma and epsilon subunits in the F-1 part and c subunit ring in the F-0 part, rotates relative to a stator composed of alpha (3)beta (3)delta ab(2) during ATP hydrolysis and synthesis. How this rotation is regulated has been less clear. Here, we show that the epsilon subunit plays a key role by acting as a switch of this motor. Two different arrangements of the E subunit have been visualized recently. The first has been observed in beef heart mitochondrial F-1-ATPase where the C-terminal portion is arranged as a two-alpha -helix hairpin structure that extends away from the alpha (3)beta (3) region, and toward the position of the c subunit ring in the intact F1F0. The second arrangement was observed in a structure determination of a complex of the gamma and E subunits of the Escherichia coli F-1-ATPase. In this, the two C-terminal helices are apart and extend along the gamma to interact with the a and P subunits in the intact complex. We have been able to trap these two arrangements by cross-linking after introducing appropriate Cys residues in E. coli F1F0. confirming that both conformations of the epsilon subunit exist in the enzyme complex. With the C-terminal domain of epsilon toward the F-0, ATP hydrolysis is activated, but the enzyme is fully coupled in both ATP hydrolysis and synthesis. With the C-terminal domain toward the F-1 part, ATP hydrolysis is inhibited and yet the enzyme is fully functional in ATP synthesis; i.e., it works in one direction only. These results help explain the inhibitory action of the epsilon subunit in the F1F0 complex and argue for a ratchet function of this subunit.
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页码:6560 / 6564
页数:5
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