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K4637 Sigma

Kinesin Protocols

  •  ISBN-10 0-89603-766-5

  •  ISBN-13 978-0-89603-766-3

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publication info   I. Vernos, ed., Humana Press, 2001, 258 pp., hard cover
mfr. no.   Humana

Description

General description

Kinesin-like proteins (KLP) are thought to be responsible for transport of cellular cargoes and involved in many cellular processes. This book presents reproducible techniques for identification, purification, and characterization of the kinesin superfamily of microtubule-dependent motors. Methods include step-by-step instructions and cautionary notes. 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.

Table of Contents

Purification of Kinesin from the Brain, Sergei A. Kuznetsov and Vladimir I. Gelfand. RT-PCR for the Identification of Developmentally Regulated Novel Members of the Kinesin-like Superfamily, Niovi Santama. Expression Cloning with Pan Kinesin Antibodies, Laura M. Ginkel and Linda Wordeman. Expression of Kinesin in Escherichia coli, Maryanne F. Stock and David D. Hackney. Plasmids for Expression of Chimeric and Truncated Kinesin Proteins, Kimberly W. Waligora and Sharyn A. Endow. Preparation of Recombinant Kinesin Superfamily Proteins Using the Baculovirus System, Nobutaka Hirokawa and Yasuko Noda. Assays for Kinesin Microtubule-Stimulated ATPase Activity, David D. Hackney and Wei Jiang. An Improved Microscope for Bead and Surface-Based Motility Assays, Nick Carter and Rob Cross. Use of Photonic Force Microscopy to Study Single-Motor-Molecule Mechanics, Sylvia Jeney, Ernst-Ludwig Florin, and J. K. Heinrich Hörber. Assays for Microtubule-Destabilizing Kinesins, Arshad Desai and Claire E. Walczak. Green Fluorescent Protein as a Tag for Molecular Motor Proteins, Sharyn A. Endow. In Vitro Reconstitution of Endosome Motility Along Microtubules, Erik Nielsen, Fedor Severin, Anthony A. Hyman, and Marino Zerial. Approaches to Study Interactions Between Kinesin Motors and Membranes, Gerardo Morfini, Ming-Ying Tsai, Györgyi Szebenyi, and Scott T. Brady. 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, and Jonathan M. Scholey. 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, and Isabelle Vernos. A Dominant Negative Approach for Functional Studies of the Kinesin II Complex, Vladimir I. Gelfand, Nathalie Le Bot, M. Carolina Tuma, and Isabelle Vernos. Identification of Kinesin-Associated Proteins, Lisa C. Lindesmith, Janardan Kumar, and Michael P. Sheetz. Assaying Spatial Organization of Microtubules by Kinesin Motors, François Nédélec and Thomas Surrey. Crystallization of Kinesin, Manfred Thormählen, Jens Müller, and Eckhard Mandelkow. Structural Analysis of the Microtubule–Kinesin Complex by Cryo-Electron Microscopy, Fabienne Beuron and Andreas Hoenger. Index.

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