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 (complimented by the forthcoming companion volume by Cassimeris and Tran) brings together current research and protocols in the field of microtoublues in vitro and will serve as a valuable tool for cell biologists, biophysicists and pharmacologists who study the microtubule cyctoskeleton, as well as for researchers in the biomedical and biotechnology communities with interest in developing drugs that target microtubules, MAPS and motors. * 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
Author(s): Leslie Wilson and John J. Correia (Eds.)
Series: Methods in Cell Biology 95
Edition: 1st
Publisher: Academic Press
Year: 2010
Language: English
Pages: 1-700
Tags: Биологические дисциплины;Цитология;
Content:
Series Editors
Page ii
Volume Editors
Page iii
Copyright
Page iv
Contributors
Pages xiii-xviii
Preface
Pages xix-xxi
Chapter 1 - Preparation of Microtubule Protein and Purified Tubulin from Bovine Brain by Cycles of Assembly and Disassembly and Phosphocellulose Chromatography
Pages 2-15
Herbert P. Miller, Leslie Wilson
Chapter 2 - Isolating Tubulin from Nonneural Sources
Pages 17-32
Dan L. Sackett, Karl A. Werbovetz, Naomi S. Morrissette
Chapter 3 - Characterization of Anti-β-tubulin Antibodies
Pages 33-46
Anthony J. Spano, Anthony Frankfurter
Chapter 4 - Expression Profiling of Tubulin Isotypes and Microtubule-Interacting Proteins Using Real-Time Polymerase Chain Reaction
Pages 47-58
Sharon Lobert, Laree Hiser, John J. Correia
Chapter 5 - Nondenaturing Electrophoresis as a Tool to Investigate Tubulin Complexes
Pages 59-75
Mónica López Fanarraga, Gerardo Carranza, Raquel Castaño, Sofia Nolasco, J. Avila, J.C. Zabala
Chapter 6 - Mass Spectrometry Analysis of C-Terminal Posttranslational Modifications of Tubulins
Pages 77-103
Virginie Redeker
Chapter 7 - Methods in Tubulin Proteomics
Pages 105-126
Leah M. Miller, Hui Xiao, Berta Burd, Susan Band Horwitz, Ruth Hogue Angeletti, Pascal Verdier-Pinard
Chapter 8 - Cryo-EM Studies of Microtubule Structural Intermediates and Kinetochore–Microtubule Interactions
Pages 128-156
Eva Nogales, Vincent H. Ramey, Hong-Wei Wang
Chapter 9 - High-Resolution Imaging of Microtubules and Cytoskeleton Structures by Atomic Force Microscopy
Pages 157-174
Loic Hamon, Patrick A. Curmi, David Pastré
Chapter 10 - Using Computational Modeling to Understand Microtubule Dynamics: A Primer for Cell Biologists
Pages 175-188
Holly V. Goodson, Ivan V. Gregoretti
Chapter 11 - Analysis of Dynamic Instability of Steady-State Microtubules In Vitro by Video-Enhanced Differential Interference Contrast Microscopy with an Appendix by Emin Oroudjev
Pages 189-206
Mythili Yenjerla, Manu Lopus, Leslie Wilson
Chapter 12 - Nanometer-Resolution Microtubule Polymerization Assays Using Optical Tweezers and Microfabricated Barriers
Pages 207-219
Blake D. Charlebois, Henry T. Schek III, Alan J. Hunt
Chapter 13 - Microtubule Dynamics Reconstituted In Vitro and Imaged by Single-Molecule Fluorescence Microscopy
Pages 221-245
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, Jonathon Howard
Chapter 14 - Studying Kinesin Motors by Optical 3D-Nanometry in Gliding Motility Assays
Pages 247-271
Bert Nitzsche, Volker Bormuth, Corina Bräuer, Jonathon Howard, Leonid Ionov, Jacob Kerssemakers, Till Korten, Cecile Leduc, Felix Ruhnow, Stefan Diez
Chapter 15 - Analysis of Tubulin Oligomers by Analytical Ultracentrifugation
Pages 274-288
John J. Correia
Chapter 16 - Determination of Drug Binding to Microtubules In Vitro
Pages 289-299
Jennifer A. Smith, Mary Ann Jordan
Chapter 17 - Fluorescence Spectroscopic Methods to Analyze Drug–Tubulin Interactions
Pages 301-329
Bhabatarak Bhattacharyya, Sonia Kapoor, Dulal Panda
Chapter 18 - A Tubulin Polymerization Microassay Used to Compare Ligand Efficacy
Pages 331-351
Ashley Davis, Sarah Martinez, Daniel Nelson, Kim Middleton
Chapter 19 - Fluorescent Taxoid Probes for Microtubule Research
Pages 353-372
Isabel Barasoain, J. Fernando Díaz, José M. Andreu
Chapter 20 - The Binding of Vinca Domain Agents to Tubulin: Structural and Biochemical Studies
Pages 373-390
Anthony Cormier, Marcel Knossow, Chunguang Wang, Benoît Gigant
Chapter 21 - Measurement of Ligand Binding to Tubulin by Sulfhydryl Reactivity
Pages 391-403
Adrian Begaye, Dan L. Sackett
Chapter 22 - Probing Interactions of Tubulin with Small Molecules, Peptides, and Protein Fragments by Solution Nuclear Magnetic Resonance
Pages 406-447
Marie-Jeanne Clément, Philippe Savarin, Elisabeth Adjadj, André Sobel, Flavio Toma, Patrick A. Curmi
Chapter 23 - Microtubule and MAPs: Thermodynamics of Complex Formation by AUC, ITC, Fluorescence, and NMR
Pages 449-480
François Devred, Pascale Barbier, Daniel Lafitte, Isabelle Landrieu, Guy Lippens, Vincent Peyrot
Chapter 24 - Quantitative Analysis of MAP-Mediated Regulation of Microtubule Dynamic Instability In Vitro: Focus on Tau
Pages 481-503
Erkan Kiris, Donovan Ventimiglia, Stuart C. Feinstein
Chapter 25 - Structure and Dynamics of the Kinesin–Microtubule Interaction Revealed by Fluorescence Polarization Microscopy
Pages 505-519
Hernando Sosa, Ana B. Asenjo, Erwin J.G. Peterman
Chapter 26 - Multiple Color Single Molecule TIRF Imaging and Tracking of MAPs and Motors
Pages 521-542
Jennifer L. Ross, Ram Dixit
Chapter 27 - Studying Plus-End Tracking at Single Molecule Resolution Using TIRF Microscopy
Pages 543-554
Ram Dixit, Jennifer L. Ross
Chapter 28 - Fluorescence Microscopy Assays on Chemically Functionalized Surfaces for Quantitative Imaging of Microtubule, Motor, and +TIP Dynamics
Pages 555-580
Peter Bieling, Ivo A. Telley, Christian Hentrich, Jacob Piehler, Thomas Surrey
Chapter 29 - Quantitative Characterization of Filament Dynamics by Single-Molecule Lifetime Measurements
Pages 582-600
Leonid A. Mirny, Daniel J. Needleman
Chapter 30 - Extracting the Mechanical Properties of Microtubules from Thermal Fluctuation Measurements on an Attached Tracer Particle
Pages 601-615
Katja M. Taute, Francesco Pampaloni, Ernst-Ludwig Florin
Chapter 31 - In Vitro Assays to Study Force Generation at Dynamic Microtubule Ends
Pages 617-639
Liedewij Laan, Marileen Dogterom
Chapter 32 - Reconstitution and Functional Analysis of Kinetochore Subcomplexes
Pages 641-656
Daniel R. Gestaut, Jeremy Cooper, Charles L. Asbury, Trisha N. Davis, Linda Wordeman
Chapter 33 - In Vitro Assays to Study the Tracking of Shortening Microtubule Ends and to Measure Associated Forces
Pages 657-676
Ekaterina L. Grishchuk, Fazly I. Ataullakhanov
Index
Pages 677-691
Volumes in Series
Pages 693-700
