PMID:11279047
| Citation |
Tanabe, M, Nishio, K, Iko, Y, Sambongi, Y, Iwamoto-Kihara, A, Wada, Y and Futai, M (2001) Rotation of a complex of the gamma subunit and c ring of Escherichia coli ATP synthase. The rotor and stator are interchangeable. J. Biol. Chem. 276:15269-74 |
|---|---|
| Abstract |
ATP synthase (F0F1) transforms an electrochemical proton gradient into chemical energy (ATP) through the rotation of a subunit assembly. It has been suggested that a complex of the gamma subunit and c ring (c(10-14)) of F0F1 could rotate together during ATP hydrolysis and synthesis (Sambongi, Y., Iko, Y., Tanabe, M., Omote, H., Iwamoto-Kihara, A., Ueda, I., Yanagida, T., Wada, Y., and Futai, M. (1999) Science 286, 1722-1724). We observed that the rotation of the c ring with the cI28T mutation (c subunit cIle-28 replaced by Thr) was less sensitive to venturicidin than that of the wild type, consistent with the antibiotic effect on the cI28T mutant and wild-type ATPase activities (Fillingame, R. H., Oldenburg, M., and Fraga, D. (1991) J. Biol. Chem. 266, 20934-20939). Furthermore, we engineered F0F1 to see the alpha(3)beta(3) hexamer rotation; a biotin tag was introduced into the alpha or beta subunit, and a His tag was introduced into the c subunit. The engineered enzymes could be purified by metal affinity chromatography and density gradient centrifugation. They were immobilized on a glass surface through the c subunit, and an actin filament was connected to the alpha or beta subunit. The filament rotated upon the addition of ATP and generated essentially the same frictional torque as one connected to the c ring. These results indicate that the gammaepsilonc(10-14) complex is a mechanical unit of the enzyme and that it can be used as a rotor or a stator experimentally, depending on the subunit immobilized. |
| Links |
PubMed Online version:10.1074/jbc.M100289200 |
| Keywords |
Actins/metabolism; Adenosine Triphosphate/metabolism; Dicyclohexylcarbodiimide/pharmacology; Enzyme Inhibitors/pharmacology; Escherichia coli/enzymology; Hydrolysis; Plasmids; Proton-Translocating ATPases/antagonists & inhibitors; Proton-Translocating ATPases/chemistry; Proton-Translocating ATPases/metabolism; Venturicidins/pharmacology |
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