title: Recombinant alpha-actin for specific fluorescent labeling.
abstract: Until recently, actin was thought to act merely as a passive track for its motility partner, myosin, during actomyosin interactions. Yet a recent report having observed dynamical conformational changes in labeled skeletal muscle alpha-actin suggests that actin has a more active role. Because the labeling technique was still immature, however, conclusions regarding the significance of the different conformations are difficult to make. Here, we describe the preparation of fully active alpha-actin obtained from a baculovirus expression system. We developed alpha-actin recombinants, of which subdomains 1 and 2 have specific sites for fluorescent probes. This specific labeling technique offers to significantly expand the information acquired from actin studies.
authors.affiliation: Nanobiology Laboratories, Graduate School of Frontier Biosciences, Osaka University, Osaka 565-0871, Japan. iwane@phys1.med.osaka-u.ac.jp
authors.forename: Atsuko H
authors.initials: AH
authors.lastname: Iwane
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Morimatsu
authors.forename: Toshio
authors.initials: T
authors.lastname: Yanagida
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D003545:Cysteine / Q000235:genetics; D005456:Fluorescent Dyes / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D017433:Protein Structure, Secondary; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000199:Actins; D005456:Fluorescent Dyes; D011994:Recombinant Proteins; D003545:Cysteine