title: In Vitro Biochemical Characterization of Cytokinesis Actin-Binding Proteins.
abstract: Characterizing the biochemical and biophysical properties of purified proteins is critical to understand the underlying molecular mechanisms that facilitate complicated cellular processes such as cytokinesis. Here we outline in vitro assays to investigate the effects of cytokinesis actin-binding proteins on actin filament dynamics and organization. We describe (1) multicolor single-molecule TIRF microscopy actin assembly assays, (2) "bulk" pyrene actin assembly/disassembly assays, and (3) "bulk" sedimentation actin filament binding and bundling assays.
authors.affiliation: Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, 60637, USA.
authors.forename: Dennis
authors.initials: D
authors.lastname: Zimmermann
authors.affiliation: Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, 60637, USA.
authors.forename: Alisha N
authors.initials: AN
authors.lastname: Morganthaler
authors.affiliation: Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, 60637, USA. drkovar@uchicago.edu.|Department of Biochemistry and Molecular Biology, The University of Chicago, 920 E. 58th St., Chicago, IL, 60637, USA. drkovar@uchicago.edu.
authors.forename: David R
authors.initials: DR
authors.lastname: Kovar
authors.affiliation: Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, 60637, USA.
authors.forename: Cristian
authors.initials: C
authors.lastname: Suarez
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D048749:Cytokinesis; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D011485:Protein Binding; D013194:Staining and Labeling
chemical_list: D000199:Actins; D002352:Carrier Proteins; D005456:Fluorescent Dyes
keywords: Actin; Biomimetics; Cytokinesis; Micropatterning; Microscopy; Single molecule; TIRF