title: Approaches to studying Arf GAPs in cells: in vitro assay with isolated focal adhesions.
abstract: The Arf GAPs are a family of proteins with a common catalytic function of hydrolyzing GTP bound to ADP-ribosylation factors (Arf) with proposed cellular functions that are diverse (Inoue and Randazzo, 2007; Kahn et al., 2008). Understanding the biochemistry of the Arf GAPs is valuable for designing and interpreting experiments using standard cell biology techniques described elsewhere. The following briefly reviews some common approaches for in vivo studies of Arf GAPs and discusses the use of isolated cellular organelles to complement in vivo experiments. Detailed protocols for examining the activity of Arf GAPs in whole cell lysates and in association with isolated focal adhesions are provided.
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland.
authors.forename: Pei-Wen
authors.initials: PW
authors.lastname: Chen
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Jian
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland.
authors.forename: Ruibai
authors.initials: R
authors.lastname: Luo
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Randazzo
mesh_terms: D020727:ADP-Ribosylation Factors / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D022001:Focal Adhesions / Q000378:metabolism* / Q000648:ultrastructure; D020690:GTPase-Activating Proteins / Q000032:analysis / Q000378:metabolism*; D006801:Humans
chemical_list: D020690:GTPase-Activating Proteins; D020727:ADP-Ribosylation Factors