Microtubules, in vitro book cover

Microtubules, in vitro

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.

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Methods in Cell Biology

Hardbound, 699 Pages

Published: June 2010

Imprint: Academic Press

ISBN: 978-0-12-374815-7


  • Section1: Isolation, Biochemistry and Characterization of Antibodies and Isotypes.

    1. nbsp;Preparation of Microtubule Protein and Purified Tubulin from Bovine Brain by Cycles of Assembly and Disassembly and Phosphocellulose Chromatography
    2. Herbert P. Miller and Leslie Wilson

    3. Measurement Of Ligand Binding To Tubulin By Sulfhydryl Reactivity
    4. Adrian Begaye and Dan L Sackett

    5. Isolating Tubulin From Nonneural Sources
    6. Dan L. Sackett, Karl A. Werbovetz and Naomi S. Morrissette

    7. Characterization of Anti-b -Tubulin Antibodies
    8. Anthony J. Spano* and Anthony Frankfurter

    9. Expression profiling of tubulin isotypes and microtubule interacting proteins using real-time polymerase chain reaction
    10. Sharon Lobert, Laree Hiser and John J. Correia

    11. Nondenaturing Electrophoresis As A Tool To Investigate Tubulin Complexes
    12. Mónica López-Fanarraga, Gerardo Carranza, Raquel Castaño, Sofia Nolasco, J. Avila and J.C. Zabala

    13. Mass spectrometry analysis of c-terminal posttranslational Modifications of tubulins
    14. Redeker, Virginie

    15. Methods in Tubulin Proteomics
    16. Leah M. Miller, Hui Xiao, Berta Burd, Susan Band Horwitz, Ruth Hogue Angeletti, Pascal Verdier-Pinard

      Section 2: Microtubule Structure and Dynamics

    17. nbsp;Cryo-EM Studies of Microtubule Structural Intermediates and Kinetochore-Microtubule Interactions
    18. Eva Nogales, Vincent H. Ramey and Hong-Wei Wang

    19. High resolution imaging of microtubules and cytoskeleton structures by Atomic Force Microscopy
    20. Loic Hamon, Patrick A. Curmi, David Pastré

    21. Holly V. Goodson and Ivan V. Gregoretti
    22. Using computational modeling to understand microtubule dynamics: a primer for cell biologists

    23. Analysis of Dynamic Instability of Steady State Microtubules In Vitro by Video-Enhanced Differential Interference Contrast Microscopy with an Appendix by Emin Oroudjev
    24. Mythili Yenjerla, Manu Lopus and Leslie Wilson

    25. Nanometer-resolution microtubule polymerization assays using optical tweezers and microfabricated barriers
    26. Blake D. Charlebois, Henry T. Schek III, Alan J. Hunt

    27. Microtubule Dynamics Reconstituted in-vitro and Imaged y Single-Molecule Fluorescence Microscopy
    28. 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

    29. Studying kinesin motors by optical 3D-nanometry in gliding motility assays
    30. Bert Nitzsche, Volker Bormuth, Corina Bräuer, Jonathon Howard, Leonid Ionov, Jacob Kerssemakers, Till Korten, Cecile Leduc, Felix Ruhnow, Stefan Diez

      Section 3: Drugs

    31. Analysis of tubulin oligomers by Analytical Ultracentrifugation.
    32. J.J. Correia

    33. nbsp;Determination of Drug Binding to Microtubules in vitro
    34. Jennifer A. Smith and Mary Ann Jordan

    35. Fluorescence Spectroscopic Methods to Analyze Drug-Tubulin Interactions
    36. Bhabatarak Bhattacharyya, Sonia Kapoor and Dulal Panda

    37. A Tubulin polymerization micro-assay used to compare ligand efficacy
    38. Ashley Davis, Sarah Martinez, Daniel Nelson and Kim Middleton.

    39. Fluorescent Taxoid Probes for Microtubule Research
    40. Isabel Barasoain, J. Fernando Díaz, and José M. Andreu

    41. nbsp;The binding of vinca domain agents to tubulin: structural and biochemical studies
    42. Anthony Cormie, Marcel Knossow, Chunguang Wang and Benoît Gigant

      Section 4: Interactions with Motors and MAPs

    43. Probing Interactions of Tubulin with Small Molecules, Peptides and Protein Fragments by Solution Nuclear Magnetic Resonance
    44. Marie-Jeanne Clément, Philippe Savarin, Elisabeth Adjadj, André Sobel, Flavio Toma, Patrick A. Curmi

    45. Microtubule and MAPs: thermodynamics of complex formation by AUC, ITC, fluorescence and NMR
    46. François Devred, Pascale Barbier, Daniel Lafitte, Isabelle Landrieu, Guy Lippens and Vincent Peyrot

    47. Quantitative Analysis of MAP Mediated Regulation of Microtubule Dynamic Instability In Vitro - Focus on Tau
    48. Erkan Kiris, Donovan Ventimiglia and Stuart C. Feinstein

    49. Structure and dynamics of the kinesin-microtubule interaction revealed by fluorescence polarization microscopy (FPM)
    50. Hernando Sosa, Ana B. Asenjo and Erwin J.G. Peterman

    51. Multiple color single molecule TIRF imaging and tracking of MAPs and motors
    52. Jennifer L. Ross and Ram Dixit

    53. Dixit and Ross - final submitted 11/4 amp; lost - resubmitted 1/8/10
    54. Fluorescence microscopy assays on chemically functionalized surfaces for quantitative imaging of microtubule, motor and +TIP dynamics
    55. Peter Bieling, Ivo A. Telley, Christian Hentrich, Jacob Piehler and Thomas Surrey

      Section 5: Functional Extracts and Force Measurements

    56. Quantitative Characterization of filament dynamics by single-molecule lifetime measurements.
    57. Leonid A. Mirny and Daniel J. Needleman

    58. Extracting the mechanical properties of microtubules from thermal fluctuation measurements on an attached tracer particle.
    59. Katja M. Taute, Francesco Pampaloni, Ernst-Ludwig Florin

    60. In vitro assays to study force generation at dynamic microtubule ends
    61. Liedewij Laan and Marileen Dogterom

    62. Reconstitution and functional analysis of kinetochore subcomplexes
    63. Daniel R. Gestaut, Jeremy Cooper, Charles L. Asbury, Trisha N. Davis and Linda Wordeman

    64. In vitro assays to study the tracking of shortening microtubule ends and to measure associated forces

    Ekaterina L. Grishchuk and Fazly I. Ataullakhanov


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