Unit

UNIT 14.12 Analysis of Arf GTP-Binding Protein Function in Cells

  1. Lee Ann Cohen,
  2. Julie G. Donaldson

Published Online: 1 SEP 2010

DOI: 10.1002/0471143030.cb1412s48

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Cohen, L. A. and Donaldson, J. G. 2010. Analysis of Arf GTP-Binding Protein Function in Cells. Current Protocols in Cell Biology. 48:14.12:14.12.1–14.12.17.

Author Information

  1. Laboratory of Cell Biology, NHLBI, National Institutes of Health, Bethesda, Maryland

Publication History

  1. Published Online: 1 SEP 2010
  2. Published Print: SEP 2010

Abstract

This unit describes techniques and approaches that can be used to study the functions of the ADP-ribosylation factor (Arf) GTP-binding proteins in cells. There are six mammalian Arfs and many more Arf-like proteins (Arls), and these proteins are conserved in eukaryotes from yeast to humans. Like all GTPases, Arfs cycle between GDP-bound, inactive and GTP-bound active conformations, facilitated by guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs) that catalyze GTP binding and hydrolysis, respectively. This unit describes approaches that can be taken to examine the localization and function of Arf and Arl proteins in cells. A simple protocol for measuring activation (GTP-binding) of specific Arf proteins in cells using a pull-down assay is also described. Approaches that can be taken to assess function of GEFs and GAPs in cells is described. Curr. Protoc. Cell Biol. 48:14.12.1-14.12.17. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • Arf;
  • GTP-binding proteins;
  • guanine nucleotide exchange factors;
  • GTPase-activating proteins