title: Purification of actin from fission yeast Schizosaccharomyces pombe and characterization of functional differences from muscle actin.
abstract: Fission yeast Schizosaccharomyces pombe is an important genetic model organism for studying the mechanisms of endocytosis and cytokinesis. However, most work on the biochemical properties of fission yeast actin-binding proteins has been done with skeletal muscle actin for matters of convenience. When simulations of mathematical models of the mechanism of endocytosis were compared with events in live cells, some of the reactions appeared to be much faster than observed in biochemical experiments with muscle actin. Here, we used gelsolin affinity chromatography to purify actin from fission yeast. S. pombe actin shares many properties with skeletal muscle actin but has higher intrinsic nucleotide exchange rate, faster trimer nucleus formation, faster phosphate dissociation rate from polymerized actin, and faster nucleation of actin filaments with Arp2/3 complex. These properties close the gap between the biochemistry and predictions made by mathematical models of endocytosis in S. pombe cells.
authors.affiliation: Department of Molecular Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8103, USA.
authors.forename: Shih-Chieh
authors.initials: SC
authors.lastname: Ti
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Pollard
mesh_terms: D051376:Actin-Related Protein 2-3 Complex / Q000737:chemistry / Q000378:metabolism; D000199:Actins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D018260:Gelsolin / Q000737:chemistry; D051379:Mice; D008954:Models, Biological; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000378:metabolism; D012568:Schizosaccharomyces / Q000737:chemistry* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D051376:Actin-Related Protein 2-3 Complex; D000199:Actins; D018260:Gelsolin; D029702:Schizosaccharomyces pombe Proteins