title: Simple in vitro assay of Arf GAPs and preparation of Arf proteins as substrates.
abstract: Defining the interaction of Arf GAPs with specific Arfs is important for understanding their functions in the endocytic system. Cell-based approaches have been valuable for identifying Arfs and Arf GAPs active in the endocytic compartment; however, the cell-based assays have some limitations in establishing relationships among the Arfs and ArfGAPs. Here we describe a simple in vitro assay that will provide a means for comparing Arfs as substrates and serve to complement cell-based studies.
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
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, MD, USA.
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, MD, USA.
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, MD, USA.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Randazzo
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D057075:Enzyme Assays; D004926:Escherichia coli; D006160:Guanosine Triphosphate / Q000737:chemistry; D006801:Humans; D006868:Hydrolysis; D053835:Unilamellar Liposomes
chemical_list: D053835:Unilamellar Liposomes; D006160:Guanosine Triphosphate; D020823:ADP-Ribosylation Factor 1
keywords: ADP-ribosylation factor; Arf GAPs; Enzyme assay; GTPase-activating protein; In vitro