GTPase Protocols: The Ras Superfamily (Methods in Molecular Biology)

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9780896039346: GTPase Protocols: The Ras Superfamily (Methods in Molecular Biology)
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In the last 10 years researchers have firmly established key roles for R- related GTPases in almost every aspect of cell biology. In the 1980s the pro- oncogene Ras itself was the focus of interest, though in the 1990s this shifted to the increasing variety of Ras-related proteins. In this new decade much yet needs to be done to establish the role for all the small GTPases now uncovered by the human genome project. In particular, these GTPases need to be und- stood in the appropriate biochemical and cellular contexts. In the process of trying to uncover the role of these versatile proteins, a variety of novel te- niques and methodologies has been developed. These now enable investi- tors to move easily within a diversity of fields ranging from structural studies to real-time in vivo analysis of a GTPase. In recognition of the need for access to key background methodologies, GTPase Protocols: The Ras Superfamily is devoted to techniques that are pr- ently widely used and that will continue to be the standard for researchers worldwide. Each chapter is aimed at supplying detailed methodologies to allow reproduction in any laboratory, while also providing the general pr- ciples on which the methods are based. Some of the techniques grouped in the first section apply broadly to small GTPases, whereas others in Part II are more applicable within each GTPase subfamily.

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Small GTPase binding proteins (GTPases) are an ancient group of proteins that play key roles in almost every aspect of cell biology, from cell proliferation to nuclear transport. In GTPase Protocols: The Ras Superfamily, Edward J. Manser and Thomas Leung have collected the key techniques currently in use to probe the function of these ubiquitous proteins both in vitro and in vivo. Presented in a format that ensures ready reproducibility by accomplished experimentalists who have refined the various methods in their laboratories, each technique includes step-by-step instructions, tips on avoiding pitfalls and troubleshooting, and ancillary notes explaining how to adapt each procedure in the event of problems. The methods cover the spectrum of core techniques required for the five major GTPase subfamilies (Ras, Rho, Rab, Arf, and Ran) and permit a diversity of applications ranging from structural studies on a GTPase to real time in vivo analysis.
Timely and highly practical, GTPase Protocols: The Ras Superfamily illuminates the powerful techniques used by investigators today to study this special family of proteins that plays such important roles in human health and disease.

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"The 20 protocols provide detailed procedures for working with most of the known small GTPases of the RAS superfamily. Included are those from RAS, RHO, RAB, ARF, and RAN. In each section, several different approaches are given, including methods for studying downstream effects. Assay methods range from microarray analysis to cell motility measurements. In each case, there is sufficient detail to allow investigators new to the procedure to use them effectively. In addition, the limitations of the procedures are provided as well as useful "notes" to facilitate adoption in the laboratory. A short bibliography accompanies each chapter and provides adequate primary source references."-Doody's Health Sciences Book Review Journal

"In GTPase Protocols, Manser and Leung assemble the key protocols required for researchers venturing into this field ... . combination of clear practical detail with underlying theory provides the reader with the information required not only to perform the technique but also to interpret the resulting data. ... In summary, this is a collection of detailed and precise experimental methods from some of the leading research groups in this field. The authors are to be congratulated on the clarity brought to each technique ... ." (Harry Mellor, Journal of Cell Science, Vol. 116 (7-8), 2003)

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