title: Isolation of Actin and Actin-Binding Proteins.
abstract: Actin-binding proteins mediate and regulate the dynamics of actin and the organization of highly ordered structures of F-actin. Villin is generally expressed in plant cells and is associated with G-actin or F-actin dependent on Ca2+concentrations. Using a DNase I affinity column chromatography approach, the villin and the G-actin can be isolated from plant material. An outline of this method including the preparation of crude protein extract from plant material, its application on the affinity column, and the successive elution of villin with a solution containing EGTA and then of G-actin with denatured reagents is presented.
authors.affiliation: Department of Life Sciences, Graduate School of Life Science, University of Hyogo, Harima Science Park City, 3-2-1 Kouto, Ako-gun, Hyogo, 678-1297, Japan. yokota@sci.u-hyogo.ac.jp.
authors.forename: Etsuo
authors.initials: E
authors.lastname: Yokota
mesh_terms: D000199:Actins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D003850:Deoxyribonuclease I / Q000737:chemistry; D027762:Lilium / Q000737:chemistry* / Q000378:metabolism; D008840:Microfilament Proteins / Q000302:isolation & purification*; D010940:Plant Proteins / Q000302:isolation & purification*; D011058:Pollen / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011489:Protein Denaturation; D020033:Protein Isoforms / Q000302:isolation & purification
chemical_list: D000199:Actins; D008840:Microfilament Proteins; D010940:Plant Proteins; D020033:Protein Isoforms; C026615:villin; D003850:Deoxyribonuclease I
keywords: Actin-binding protein; DNase I affinity column chromatography; F-Actin; G-actin; Villin