Identification of an uncoupling mutation affecting the b subunit of F1F0 ATP synthase in Escherichia coli

A specific b subunit arginine, b(Arg-36) in Escherichia coli, displays evolutionary conservation among bacterial F1F0 ATP synthases. Site-directed mutagenesis was used to generate a collection of mutations affecting b(Arg-36). The phenotype differed depending upon the substitution, and the b(Arg-36-->Glu) and b(Arg-36-->Ile) substitutions virtually abolished enzyme, function. Although the total amounts of F1F0 ATP synthase present in the membranes prepared from mutant strains were reduced, the primary effect of the b(Arg-36) substitutions was on the activities of the intact enzyme complexes. The most interesting result was that the b(Arg-36-->Glu) substitution results in the uncoupling of a functional F-0 from F-1 ATP hydrolysis activity. (C) 1998 Federation of European Biochemical Societies.