F(o)F1-ATP synthase mediates coupling of proton flow in F(o) and ATP synthesis/hydrolysis in F1 through rotation of central rotor subunits. A ring structure of F(o)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(o)c subunit ring in detergent-solubilized F(o)F1-ATP synthase purified from Escherichia coli. Similar studies have been performed and reported recently [Sambongi et 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(o) inhibitor. We have found that detergents which adequately disperse the enzyme for the rotation assay also tend to transform F(o)F1-ATP synthase into an F(o) inhibitor-insensitive state in which F1 can hydrolyze ATP regardless of the state of the F(o). Our results raise the important issue of whether rotation of the F(o)c ring in isolated F(o)F1-ATP synthase can be demonstrated unequivocally with the approach adopted here and also used by Sambongi et al. Copyright (C) 2000 Federation of European Biochemical Societies.
ASJC Scopus subject areas
- Molecular Biology