General Description of the Volume: 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. General Description of the Series: The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences.

Key Features

@introbul:Key Features @bul:* Assays to measure GTP exchange and hydrolysis * Expression and purification of small GTPases * Posttranslational modifications of Rab proteins * Generation of trans dominant mutants * Expression and purification of Rab effector proteins


Biochemists, molecular biologists, cell biologists, pharmacologists, neurophysiologists, and neurochemists.

Table of Contents

Expression, Purification, and Posttranslational Modification: C. Nuoffer, F. Peter, and W.E. Balch, Purification of His6-Tagged Rab1 Proteins Using Bacterial and Insect Cell Expression Systems. H. Horiuchi,O. Ullrich, C. Bucci, and M. Zerial, Purification of Posttranslationally Modified and Unmodified Rab5 Protein Expressed in Spodoptera frugiperda Cells. M.A. Riederer, T. Soldati, A.B. Dirac-Svejstrup, and S.R. Pfeffer, Expression of Rab9 Protein in Escherichia coli: Purification and Isoprenylation in Vitro. Y. Jiang, G. Rossi, and S. Ferro-Novick, Characterization of Yeast Type-II Geranylgeranyltransferase. S.A. Armstrong, M.S. Brown, J.L. Goldstein, and M.C. Seabra, Preparation of Recombinant Rab Geranylgeranyltransferase and Rab Escort Proteins. K. Kimura, T. Oka, and A. Nakano, Purification and Assay of Yeast Sar1p. T. Rowe and W.E. Balch, Expression and Purification of Mammalian Sar1. Guanine NucleotideExchange and Hydrolysis: A. Kikuchi, H. Nakanishi, and Y. Takai, Purification and Properties of Rab3A. T. Sasaki and Y. Takai, Purification and Properties of Bovine Rab-GDP Dissociation Inhibitor. F. Peter, C. Nuoffer, I. Schalk, and W.E. Balch, Expression and Purification of Recombinant His6-Tagged Guanine Nucleotide Dissociation Inhibitor and Formation of Rab1 Complex. D.M. Roberts, M.D. Garrett, and P.J. Novick, Purification of GDP Dissociation Stimulator Dss4 from Recombinant Bacteria. J.L. Burton and P. De Camilli, Expression, Purification, and Functional Assay of Mss4. C. Barlowe and R. Schekman, Expression, Purification, and Assay of Sec12p: A Sar1p-Specific GDP Dissociation Stimulator.


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© 1995
Academic Press
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@from:Praise for the Series @qu:"The Methods in Enzymology series represents the gold-standard." @source:--NEUROSCIENCE @qu:"Incomparably useful." @source:--ANALYTICAL BIOCHEMISTRY @qu:"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." @source:--BIO/TECHNOLOGY @qu:"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." @source:--CHEMISTRY IN INDUSTRY @qu:"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." @source:--AMERICAN SOCIETY OF MICROBIOLOGY NEWS @qu:"If we had some way to find the work most often consulted in the laboratory, it could well be the multi-volume series Methods in Enzymology...a great work." @source:--ENZYMOLOGIA @qu:"A series that has established itself as a definitive reference for biochemists." @source:--JOURNAL OF CHROMATOGRAPHY