Microtubules, in vitro

Microtubules, in vitro

1st Edition - June 1, 2010

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  • Editors: John Correia, Leslie Wilson
  • eBook ISBN: 9780080888583

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Description

There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that control cell cycle, dynamics, organization and transport. The field continues to grow and explore new aspects of these issues driven immensely by developments in optical imaging and tracking techniques. This volume (complemented by the forthcoming companion volume by Cassimeris and Tran) brings together current research and protocols in the field of microtoubules in vitro and will serve as a valuable tool for cell biologists, biophysicists and pharmacologists who study the microtubule cytoskeleton, as well as for researchers in the biomedical and biotechnology communities with interest in developing drugs that target microtubules, MAPS and motors.

Key Features

  • Chapters reflect both experimental procedures and new developments in the field of microtubule in vitro research
  • Combines classical approaches and modern technologies
  • Presents easy-to-use protocols and thorough background information, compiled by leaders in the field

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Table of Contents

  • Section1: Isolation, Biochemistry and Characterization of Antibodies and Isotypes
    1. Preparation of Microtubule Protein and Purified Tubulin from Bovine Brain by Cycles of Assembly and Disassembly and Phosphocellulose Chromatography
    Herbert P. Miller and Leslie Wilson
    2. Measurement Of Ligand Binding To Tubulin By Sulfhydryl Reactivity
    Adrian Begaye and Dan L Sackett
    3. Isolating Tubulin From Nonneural Sources
    Dan L. Sackett, Karl A. Werbovetz and Naomi S. Morrissette
    4. Characterization of Anti-b -Tubulin Antibodies
    Anthony J. Spano* and Anthony Frankfurter
    5. Expression profiling of tubulin isotypes and microtubule interacting proteins using real-time polymerase chain reaction
    Sharon Lobert, Laree Hiser and John J. Correia
    6. Nondenaturing Electrophoresis As A Tool To Investigate Tubulin Complexes
    Mónica López-Fanarraga, Gerardo Carranza, Raquel Castaño, Sofia Nolasco, J. Avila and J.C. Zabala
    7 .Mass spectrometry analysis of c-terminal posttranslational Modifications of tubulins
    Redeker, Virginie
    8. Methods in Tubulin Proteomics
    Leah M. Miller, Hui Xiao, Berta Burd, Susan Band Horwitz, Ruth Hogue Angeletti, Pascal Verdier-Pinard

    Section 2: Microtubule Structure and Dynamics
    9. Cryo-EM Studies of Microtubule Structural Intermediates and Kinetochore-Microtubule Interactions
    Eva Nogales, Vincent H. Ramey and Hong-Wei Wang
    10. High resolution imaging of microtubules and cytoskeleton structures by Atomic Force Microscopy
    Loic Hamon, Patrick A. Curmi, David Pastré
    11. Using computational modeling to understand microtubule dynamics: a primer for cell biologists
    Holly V. Goodson and Ivan V. Gregoretti
    12. Analysis of Dynamic Instability of Steady State Microtubules In Vitro by Video-Enhanced Differential Interference Contrast Microscopy with an Appendix by Emin Oroudjev
    Mythili Yenjerla, Manu Lopus and Leslie Wilson
    13. Nanometer-resolution microtubule polymerization assays using optical tweezers and microfabricated barriers
    Blake D. Charlebois, Henry T. Schek III, Alan J. Hunt
    14. Microtubule Dynamics Reconstituted in-vitro and Imaged y Single-Molecule Fluorescence Microscopy
    Christopher Gell, Volker Bormuth, Gary J. Brouhard, Daniel N. Cohen, Stefan Diez, Claire T. Friel, Jonne Helenius, Bert Nitzsche, Heike Petzold, Jan Ribbe, Erik Schäffer, Jeffrey H. Stear, Anastasiya Trushko, Vladimir Varga, Per O. Widlund, Marija Zanic, and Jonathon Howard
    15. Studying kinesin motors by optical 3D-nanometry in gliding motility assays
    Bert Nitzsche, Volker Bormuth, Corina Bräuer, Jonathon Howard, Leonid Ionov, Jacob Kerssemakers, Till Korten, Cecile Leduc, Felix Ruhnow, Stefan Diez

    Section 3: Drugs
    16. Analysis of tubulin oligomers by Analytical Ultracentrifugation.
    J.J. Correia
    17. Determination of Drug Binding to Microtubules in vitro
    Jennifer A. Smith and Mary Ann Jordan
    18. Fluorescence Spectroscopic Methods to Analyze Drug-Tubulin Interactions
    Bhabatarak Bhattacharyya, Sonia Kapoor and Dulal Panda
    19. A Tubulin polymerization micro-assay used to compare ligand efficacy
    Ashley Davis, Sarah Martinez, Daniel Nelson and Kim Middleton.
    20. Fluorescent Taxoid Probes for Microtubule Research
    Isabel Barasoain, J. Fernando Díaz, and José M. Andreu
    21. The binding of vinca domain agents to tubulin: structural and biochemical studies
    Anthony Cormie, Marcel Knossow, Chunguang Wang and Benoît Gigant

    Section 4: Interactions with Motors and MAPs
    22. Probing Interactions of Tubulin with Small Molecules, Peptides and Protein Fragments by Solution Nuclear Magnetic Resonance
    Marie-Jeanne Clément, Philippe Savarin, Elisabeth Adjadj, André Sobel, Flavio Toma, Patrick A. Curmi
    23. Microtubule and MAPs: thermodynamics of complex formation by AUC, ITC, fluorescence and NMR
    François Devred, Pascale Barbier, Daniel Lafitte, Isabelle Landrieu, Guy Lippens and Vincent Peyrot
    24. Quantitative Analysis of MAP Mediated Regulation of Microtubule Dynamic Instability In Vitro – Focus on Tau
    Erkan Kiris, Donovan Ventimiglia and Stuart C. Feinstein
    25. Structure and dynamics of the kinesin-microtubule interaction revealed by fluorescence polarization microscopy (FPM)
    Hernando Sosa, Ana B. Asenjo and Erwin J.G. Peterman
    26. Multiple color single molecule TIRF imaging and tracking of MAPs and motors
    Jennifer L. Ross and Ram Dixit
    27. Fluorescence microscopy assays on chemically functionalized surfaces for quantitative imaging of microtubule, motor and +TIP dynamics
    Peter Bieling, Ivo A. Telley, Christian Hentrich, Jacob Piehler and Thomas Surrey

    Section 5: Functional Extracts and Force Measurements
    28. Quantitative Characterization of filament dynamics by single-molecule lifetime measurements.
    Leonid A. Mirny and Daniel J. Needleman
    29. Extracting the mechanical properties of microtubules from thermal fluctuation measurements on an attached tracer particle. =
    Katja M. Taute, Francesco Pampaloni, Ernst-Ludwig Florin
    30. In vitro assays to study force generation at dynamic microtubule ends
    Liedewij Laan and Marileen Dogterom
    31. Reconstitution and functional analysis of kinetochore subcomplexes
    Daniel R. Gestaut, Jeremy Cooper, Charles L. Asbury, Trisha N. Davis and Linda Wordeman
    32. In vitro assays to study the tracking of shortening microtubule ends and to measure associated forces
    Ekaterina L. Grishchuk and Fazly I. Ataullakhanov

Product details

  • No. of pages: 699
  • Language: English
  • Copyright: © Academic Press 2010
  • Published: June 1, 2010
  • Imprint: Academic Press
  • eBook ISBN: 9780080888583

About the Editors

John Correia

Affiliations and Expertise

University of Mississippi Medical Center, Jackson, USA

Leslie Wilson

University of California, Santa Barbara, USA

Affiliations and Expertise

University of California, Santa Barbara, USA

About the Serial Volume Editors

Leslie Wilson

University of California, Santa Barbara, USA

Affiliations and Expertise

University of California, Santa Barbara, USA

John Correia

Affiliations and Expertise

University of Mississippi Medical Center, Jackson, USA

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