UNIT 14.12 Analysis of Arf GTP-Binding Protein Function in Cells
Published Online: 1 SEP 2010
Copyright © 2010 by John Wiley & Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 SEP 2010
- Published Print: SEP 2010
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.
- GTP-binding proteins;
- guanine nucleotide exchange factors;
- GTPase-activating proteins