title: Subunit rotation of vacuolar-type proton pumping ATPase: relative rotation of the G and C subunits.
abstract: Vacuolar-type ATPases V1V0 (V-ATPases) are found ubiquitously in the endomembrane organelles of eukaryotic cells. In this study, we genetically introduced a His tag and a biotin tag onto the c and G subunits, respectively, of Saccharomyces cerevisiae V-ATPase. Using this engineered enzyme, we observed directly the continuous counter-clockwise rotation of an actin filament attached to the G subunit when the enzyme was immobilized on a glass surface through the c subunit. V-ATPase generated essentially the same torque as the F-ATPase (ATP synthase). The rotation was inhibited by concanamycin and nitrate but not by azide. These results demonstrated that the V- and F-ATPase carry out a common rotational catalysis.
authors.affiliation: Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Hirata
authors.forename: Atsuko
authors.initials: A
authors.lastname: Iwamoto-Kihara
authors.forename: Ge-Hong
authors.initials: GH
authors.lastname: Sun-Wada
authors.forename: Toshihide
authors.initials: T
authors.lastname: Okajima
authors.forename: Yoh
authors.initials: Y
authors.lastname: Wada
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Futai
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry; D001710:Biotin; D006639:Histidine; D020409:Molecular Motor Proteins / Q000737:chemistry*; D015202:Protein Engineering; D021122:Protein Subunits / Q000737:chemistry; D006180:Proton-Translocating ATPases; D012399:Rotation; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D017385:Sequence Homology; D025262:Vacuolar Proton-Translocating ATPases / Q000737:chemistry*
chemical_list: D020409:Molecular Motor Proteins; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D006639:Histidine; D001710:Biotin; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases