title: Charge-reversion mutagenesis of Dictyostelium actin to map the surface recognized by myosin during ATP-driven sliding motion.
abstract: Amino acid residues D24/D25, E99/E100, E360/E361, and D363/E364 in subdomain 1 of Dictyostelium actin were replaced with histidine residues by site-directed mutagenesis. Mutant actins were expressed in Dictyostelium cells and purified to homogeneity. The sliding movement of mutant actin filaments on heavy meromyosin attached to a glass surface was measured to assess the effect of the mutation on the motility of actin. For two C-terminal mutants, force generated by a single actin filament and myosin was also measured. These measurements indicated that both D24/D25 and E99/E100 are involved in ATP-driven sliding, whereas E360/E361/D363/E364 are not essential for ATP-driven sliding and force generation.
authors.affiliation: Department of Pure and Applied Sciences, College of Arts and Sciences, University of Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Johara
authors.forename: Y Y
authors.initials: YY
authors.lastname: Toyoshima
authors.forename: A
authors.initials: A
authors.lastname: Ishijima
authors.forename: H
authors.initials: H
authors.lastname: Kojima
authors.forename: T
authors.initials: T
authors.lastname: Yanagida
authors.forename: K
authors.initials: K
authors.lastname: Sutoh
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D004023:Dictyostelium / Q000235:genetics / Q000378:metabolism*; D006639:Histidine; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D015879:Myosin Subfragments / Q000378:metabolism; D009218:Myosins / Q000378:metabolism*; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D000199:Actins; D015879:Myosin Subfragments; D011994:Recombinant Proteins; D006639:Histidine; D000255:Adenosine Triphosphate; D009218:Myosins