E-Book Overview
Small GTPases play a key role in many aspects of contemporary cell biology: control of cell growth and differentiation; regulation of cell adhesion and cell movement; the organization of the actin cytoskeleton; and the regulation of intracellular vesicular transport. This volume plus its companion Volumes 255 and 256 cover all biochemical and biological assays currently in use for analyzing the role of small GTPases in these aspects of cell biology at the molecular level.Volume 257 provides detailed protocols to effectively produce, modify, and assay for the function of small GTPases involved in vesicular traffic through the secretory pathway of eukaryotic cells.
E-Book Content
Preface GTPases are now recognized as essential components for protein traffic between all compartments of the cell. This includes vesicular traffic through the exocytic and endocytic pathways, where GTPases play key roles in the assembly of vesicle coats (budding), in vesicle targeting and in fusion, as well as in protein traffic in and out of the nucleus. GTPases involved in transport include the Rab and A R F families, Sarl, Ran, dynamin, and heterotrimeric G proteins. In addition to GTPase, a number of associated accessory factors are critical for function. These include posttranslational modifying enzymes (such as prenyl transferases and myristyl transferases), factors that affect guanine nucleotide binding [guanine nucleotide dissociation inhibitors (GDIs) and guanine nucleotide exchange factors (GEFs)], and factors that stimulate guanine nucleotide hydrolysis [GTPase-activating proteins (GAPs)]. To understand the function of GTPases and their cognate factors, a wealth of in vitro biochemical and in vivo molecular genetic approaches are currently being applied to individual proteins. Given the diverse spectrum of compartments regulated by individual GTPases, techniques developed for one particu