title: A mutation in which alanine 128 Is replaced by aspartic acid abolishes dimerization of the b-subunit of the F0F1-ATPase from Escherichia coli.
abstract: Site-directed mutagenesis was used to investigate the roles of a short series of hydrophobic amino acids in the b-subunit of the Escherichia coli F0F1-ATPase. A mutation affecting one of these, G131D, had been previously characterized and was found to interrupt assembly of the F0F1-ATPase (Jans, D. A., Hatch, L., Fimmel, A. L., Gibson, D., and Cox, G. B. (1985) J. Bacteriol. 162, 420-426). To extend this work, aspartic acid was substituted for each one of the residues from positions 124 to 132. The properties of mutants in this series are consistent with the region from Val124 to Gly131 forming an alpha-helix. Two of the mutations, V124D and A128D, resulted in a similar phenotype to the G131D mutation. This suggested that Val124, Ala128, and Gly131 form a helical face which may have a role in inter- or intrasubunit interactions. This was tested by overexpressing and purifying the cytoplasmic domains of the wild type and A128D mutant b-subunits. Sedimentation equilibrium centrifugation indicated that the wild type domain formed a dimer whereas the mutant was present as a monomer.
authors.affiliation: Division of Biochemistry and Molecular Biology, John Curtin School of Medical Research, Australian National University, GPO Box 334, Canberra City, ACT 2601 Australia.
authors.forename: S M
authors.initials: SM
authors.lastname: Howitt
authors.forename: A J
authors.initials: AJ
authors.lastname: Rodgers
authors.forename: P D
authors.initials: PD
authors.lastname: Jeffrey
authors.forename: G B
authors.initials: GB
authors.lastname: Cox
mesh_terms: D000409:Alanine / Q000235:genetics*; D001224:Aspartic Acid / Q000235:genetics*; D001704:Biopolymers; D004926:Escherichia coli / Q000201:enzymology*; D016297:Mutagenesis, Site-Directed; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics*
chemical_list: D001704:Biopolymers; D001224:Aspartic Acid; D006180:Proton-Translocating ATPases; D000409:Alanine