title: Stepping rotation of F(1)-ATPase with one, two, or three altered catalytic sites that bind ATP only slowly.
abstract: F(1)-ATPase is an ATP hydrolysis-driven motor in which the gamma subunit rotates in the stator cylinder alpha(3)beta(3). To know the coordination of three catalytic beta subunits during catalysis, hybrid F(1)-ATPases, each containing one, two, or three "slow" mutant beta subunits that bind ATP very slowly, were prepared, and the rotations were observed with a single molecule level. Each hybrid made one, two, or three steps per 360 degrees revolution, respectively, at 5 microm ATP where the wild-type enzyme rotated continuously without step under the same observing conditions. The observed dwell times of the steps are explained by the slow binding rate of ATP. Except for the steps, properties of rotation, such as the torque forces exerted during rotary movement, were not significantly changed from those of the wild-type enzyme. Thus, it appears that the presence of the slow beta subunit(s) does not seriously affect other normal beta subunit(s) in the same F(1)-ATPase molecule and that the order of sequential catalytic events is faithfully maintained even when ATP binding to one or two of the catalytic sites is retarded.
authors.affiliation: Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226-8503, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ariga
authors.forename: Tomoko
authors.initials: T
authors.lastname: Masaike
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Noji
authors.forename: Masasuke
authors.initials: M
authors.lastname: Yoshida
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D001483:Base Sequence; D020134:Catalytic Domain; D017931:DNA Primers; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases