Observations of rotation within the F_OF_l-ATP synthase:: deciding between rotation of the F_O<i>c</i> subunit ring and artifact

作     者：Tsunoda, SP Aggeler, R Noji, H Kinosita, K Yoshida, M Capaldi, RA 

作者机构：Tokyo Inst Technol Resources Utilizat Res Lab Yokohama Kanagawa 2268503 Japan Univ Oregon Inst Mol Biol Eugene OR 97403 USA Teikyo Univ Biotechnol Res Ctr 3F CREST Genet Programming Team 13 Kawasaki Kanagawa 2160001 Japan Keio Univ Fac Sci & Technol Dept Phys Yokohama Kanagawa 2238522 Japan 

出 版 物：《FEBS LETTERS》 (FEBS Lett.)

年 卷 期：2000年第470卷第3期

页      面：244-248页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 07[理学] 

基　　金：Japan Society for the Promotion of Science  KAKEN 

主　　题：FoFl-ATP synthase rotation c subunit ring detergent uncoupling 

摘      要：F0F1-ATP synthase mediates coupling of proton flow in F-0 and ATP synthesis/hydrolysis in F(0)c through rotation of central rotor subunits. A ring structure of F(0)c subunits is widely believed to be a part of the rotor. Using an attached actin filament as a probe, we have observed the rotation of the F(0)c subunit ring in detergent-solubilized F0F1-ATP synthase purified from Escherichia coli. Similar studies have been performed and reported recently [Sambongi ct al, (1999) Science 286, 1722-1724], However. in our hands this rotation has been observed only for the preparations which show poor sensitivity to dicyclohexylcarbodiimde, an F-0 inhibitor. We hare found that detergents which adequately disperse the enzyme for the rotation assay also tend to transform F0F1-ATP synthase into an F-0 inhibitor-insensitive state in which Fl can hydrolyze ATP regardless of the state of the F-0. Our results raise the important issue of whether rotation of the F(0)c ring in isolated F0F1-ATP synthase can be demonstrated unequivocally with the approach adopted here and also used by Sambongi et al. :(C) 2000 Federation of European Biochemical Societies.