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.

Keywords: ADP-ribosylation factor; Arf GAPs; Enzyme assay; GTPase-activating protein; In vitro.