Kinesin protocols / edited by Isabelle Vernos.

Format:

Book

Contributor:

Vernos, Isabelle.

Series:

Methods in molecular biology (Clifton, N.J.) ; v. 164.

Methods in molecular biology ; v. 164

Language:

English

Subjects (All):

Kinesin--Laboratory manuals.

Kinesin.

Kinesins.

Medical Subjects:

Kinesins.

Genre:

Laboratory manuals.

Physical Description:

xi, 258 pages : illustrations ; 24 cm.

Place of Publication:

Totowa, NJ : Humana Press, 2001.

Summary:

Isabelle Vernos and a panel of hands-on experts present their most productive and reproducible techniques for the identification, purification, and characterization of the kinesin superfamily of microtubule-dependent motors. The methods range from the most basic to the most sophisticated and include step-by-step instructions and extensive cautionary notes to ensure experimental success. Among the approaches considered are methods to express and purify kinesins in different systems, to characterize microtubule-enhanced ATPase activity and motility properties, and to test microtubule destabilizing activity. Comprehensive and highly practical, Kinesin Protocols makes available all the key basic and cutting-edge methods needed to successfully study the multifaceted world of kinesin-like proteins and to explore their many functions.

Contents:

1 Purification of Kinesin from the Brain / Sergei A. Kuznetsov, Vladimir I. Gelfand 1

2 RT-PCR for the Identification of Developmentally Regulated Novel Members of the Kinesin-like Superfamily / Niovi Santama 9

3 Expression Cloning with Pan Kinesin Antibodies / Laura M. Ginkel, Linda Wordeman 21

4 Expression of Kinesin in Escherichia coli / Maryanne F. Stock, David D. Hackney 43

5 Plasmids for Expression of Chimeric and Truncated Kinesin Proteins / Kimberly W. Waligora, Sharyn A. Endow 49

6 Preparation of Recombinant Kinesin Superfamily Proteins Using the Baculovirus System / Nobutaka Hirokawa, Yasuko Noda 57

7 Assays for Kinesin Microtubule-Stimulated ATPase Activity / David D. Hackney, Wei Jiang 65

8 An Improved Microscope for Bead and Surface-Based Motility Assays / Nick Carter, Rob Cross 73

9 Use of Photonic Force Microscopy to Study Single-Motor-Molecule Mechanics / Sylvia Jeney, Ernst-Ludwig Florin, J. K. Heinrich Horber 91

10 Assays for Microtubule-Destabilizing Kinesins / Arshad Desai, Claire E. Walczak 109

11 Green Fluorescent Protein as a Tag for Molecular Motor Proteins / Sharyn A. Endow 123

12 In Vitro Reconstitution of Endosome Motility Along Microtubules / Erik Nielsen, Fedor Severin, Anthony A. Hyman, Marino Zerial 133

13 Approaches to Study Interactions Between Kinesin Motors and Membranes / Gerardo Morfini, Ming-Ying Tsai, Gyorgyi Szebenyi, Scott T. Brady 147

14 Microinjection Methods for Analyzing the Functions of Kinesins in Early Embryos / Robert L. Morris, Heather M. Brown, Brent D. Wright, David J. Sharp, William Sullivan, Jonathan M. Scholey 163

15 The Use of Dominant Negative Mutants to Study the Function of Mitotic Motors in the In Vitro Spindle Assembly Assay in Xenopus Egg Extracts / Haralabia Boleti, Eric Karsenti, Isabelle Vernos 173

16 A Dominant Negative Approach for Functional Studies of the Kinesin II Complex / Vladimir I. Gelfand, Nathalie Le Bot, M. Carolina Tuma, Isabelle Vernos 191

17 Identification of Kinesin-Associated Proteins / Lisa C. Lindesmith, Janardan Kumar, Michael P. Sheetz 205

18 Assaying Spatial Organization of Microtubules by Kinesin Motors / Francois Nedelec, Thomas Surrey 213

19 Crystallization of Kinesin / Manfred Thormahlen, Jens Muller, Eckhard Mandelkow 223

20 Structural Analysis of the Microtubule-Kinesin Complex by Cryo-Electron Microscopy / Fabienne Beuron, Andreas Hoenger 235.

Notes:

Includes bibliographical references and index.

ISBN:

0896037665

OCLC:

43845649