title: Purification and characterization of a Ca2+-dependent actin filament severing protein from bovine adrenal medulla.
abstract: We describe the purification of an actin regulatory protein from bovine adrenal medulla. This protein caused a dose-dependent decrease of the specific viscosity of actin solution within 30 s of its addition in a Ca2+-sensitive way. Sedimentation assays and the observation by electron microscopy showed that this effect was ascribable to the fragmentation of actin filaments. This protein apparently promoted nucleation of actin polymerization and increased the critical concentration of actin for polymerization nearly 5-fold, suggesting its binding to the barbed end of actin filaments. The inhibitory effect of this protein on the elongation of actin from the barbed end of the myosin subfragment S1-labeled actin seeds confirmed this suggestion. These properties are similar to those of gelsolin. However, the physicochemical properties of this protein having a single polypeptide chain with a molecular weight of 74,000, a Stokes radius of 3.9 nm, a sedimentation coefficient (s0(20),w) of 4.5 S, and an immunological characterization showed that this protein is different from gelsolin.
authors.affiliation: Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Maekawa
authors.forename: M
authors.initials: M
authors.lastname: Toriyama
authors.forename: S
authors.initials: S
authors.lastname: Hisanaga
authors.forename: N
authors.initials: N
authors.lastname: Yonezawa
authors.forename: S
authors.initials: S
authors.lastname: Endo
authors.forename: N
authors.initials: N
authors.lastname: Hirokawa
authors.forename: H
authors.initials: H
authors.lastname: Sakai
mesh_terms: D008841:Actin Cytoskeleton / Q000648:ultrastructure; D000199:Actins / Q000378:metabolism*; D000313:Adrenal Medulla / Q000032:analysis*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000502:physiology*; D002417:Cattle; D008840:Microfilament Proteins / Q000302:isolation & purification* / Q000378:metabolism; D008854:Microscopy, Electron; D008970:Molecular Weight; D010449:Peptide Mapping; D011108:Polymers; D011485:Protein Binding; D014783:Viscosity
chemical_list: D000199:Actins; D000596:Amino Acids; D008840:Microfilament Proteins; D011108:Polymers; D002118:Calcium