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GTPase Protocols

The Ras Superfamily

  • Book
  • © 2002

Overview

Editors:
  1. Ed Manser

    1. Glaxo-IMCB Group, Institute of Molecular and Cell Biology, Singapore

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    You can also search for this editor in PubMed   Google Scholar

  2. Thomas Leung

    1. Glaxo-IMCB Group, Institute of Molecular and Cell Biology, Singapore

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

Part of the book series: Methods in Molecular Biology (MIMB, volume 189)

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Table of contents (20 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. General Protocols

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. The GTPase Cycle How Dominant Inhibitory Mutants Block the Biological Functions of Small GTPases

      • Edward J. Manser
      Pages 3-11

    3. Preparation of GTPases for Structural and Biophysical Analysis

      • Susan J. M. Smith, Katrin Rittinger
      Pages 13-24

    4. Using cDNA-Representational Difference Analysis (cDNA-RDA) in Combination with Microarrays to Identify Rac Regulated Genes

      • Arndt A. P. Schmitz, Robert Lucito, Linda Van Aelst
      Pages 25-43

    5. Fluorescence Methods in the Study of Small GTP-Binding Proteins

      • Mohammad Reza Ahmadian, Alfred Wittinghofer, Christian Herrmann
      Pages 45-63

  3. Specific Methods

    1. Front Matter

      Pages 65-65
      Download chapter PDF

    2. Ras and Rac as Activators of Reactive Oxygen Species (ROS)

      • Herbert Archer, Dafna Bar-Sagi
      Pages 67-73

    3. Interfering with Ras Signaling Using Membrane-Permeable Peptides or Drugs

      • Hiroshi Maruta, Hong He, Thao Nheu
      Pages 75-86

    4. A Ras-Based Module to Generate 32P-Labeled Fusion Proteins for Blot Overlays

      • Zhoushen Zhao, Louis Lim
      Pages 87-95

    5. Determination of the Activity of Rho-Like GTPases in Cells

      • Angeliki Malliri, Jean P. Ten Klooster, Cristina Olivo, John G. Collard
      Pages 99-109

    6. Introduction of Dominant Inhibitory Proteins Directed Against ROK and MRCK Kinases

      • Ivan Tan, Louis Lim, Thomas Leung
      Pages 111-119

    7. Effects of Rho Family GTPases on Cell-Cell Adhesion

      • Masaki Fukata, Masato Nakagawa, Shinya Kuroda, Kozo Kaibuchi
      Pages 121-128

    8. Cell Motility and Invasion Assays

      • Jackie Banyard, Marc Symons
      Pages 129-140

    9. Isolation of Regulatory Proteins for the Rab3 Subfamily GTPases

      • Hiroyuki Nakanishi, Takuya Sasaki, Yoshimi Takai
      Pages 143-156

    10. Analysis and Preparation of Stable Complexes between Rab GTPases, Rab Escort Protein, and Rab Geranylgeranyl Transferase

      • José B. Pereira-Leal, Anita Q. Gomes, Miguel C. Seabra
      Pages 157-165

    11. Preparation of Myristoylated Arf1 and Arf6 Proteins

      • Paul A. Randazzo, Henry M. Fales
      Pages 169-179

    12. ARF-Directed Guanine-Nucleotide-Exchange (GEP) Proteins

      • Gustavo Pacheco-Rodriguez, Joel Moss, Martha Vaughan
      Pages 181-189

    13. Arf6 and its Role in Cytoskeletal Modulation

      • Julie G. Donaldson
      Pages 191-198

    14. ARF GTPase-Activating Protein 1

      • Irit Huber, Edna Cukierman, Miriam Rotman, Dan Cassel
      Pages 199-206
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About this book

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.

Reviews

From the reviews:

"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)

Editors and Affiliations

  • Glaxo-IMCB Group, Institute of Molecular and Cell Biology, Singapore

    Ed Manser, Thomas Leung

Bibliographic Information

  • Book Title: GTPase Protocols

  • Book Subtitle: The Ras Superfamily

  • Editors: Ed Manser, Thomas Leung

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/1592592813

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2002

  • Hardcover ISBN: 978-0-89603-934-6Published: 05 June 2002

  • Softcover ISBN: 978-1-61737-261-2Published: 10 November 2010

  • eBook ISBN: 978-1-59259-281-4Published: 03 February 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 267

  • Topics: Biochemistry, general

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