Microtubules, in vitro

Microtubules, in vitro

2nd Edition - August 20, 2013
  • Editors: John Correia, Leslie Wilson
  • Hardcover ISBN: 9780124077577
  • eBook ISBN: 9780124078888

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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 2e brings together current research and protocols in the field of microtubules 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 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

Readership

Researchers and students in cell, molecular and developmental biology

Table of Contents

  • Series Page

    Contributors

    Preface

    Chapter 1. Site-Specific Fluorescent Labeling of Tubulin

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 2. Measuring Microtubule Persistence Length Using a Microtubule Gliding Assay

    Abstract

    Introduction

    Summary

    Acknowledgments

    References

    Chapter 3. Structural Studies of the Doublecortin Family of MAPs

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 4. Detection and Quantification of Microtubule Detachment from Centrosomes and Spindle Poles

    Abstract

    Introduction

    Summary

    Acknowledgments

    References

    Chapter 5. Regulation of Tubulin Expression by Micro-RNAs: Implications for Drug Resistance

    Abstract

    Introduction and Rationale

    Summary

    References

    Chapter 6. Determining the Structure–Mechanics Relationships of Dense Microtubule Networks with Confocal Microscopy and Magnetic Tweezers-Based Microrheology

    Abstract

    Introduction

    Summary

    Acknowledgments

    References

    Chapter 7. Studying Mitochondria and Microtubule Localization and Dynamics in Standardized Cell Shapes

    Abstract

    Introduction

    Conclusion

    Acknowledgments

    References

    Chapter 8. Going Solo: Measuring the Motions of Microtubules with an In Vitro Assay for TIRF Microscopy

    Abstract

    Introduction

    Concluding Remarks

    Acknowledgments

    References

    Chapter 9. Analysis of Microtubules in Isolated Axoplasm from the Squid Giant Axon

    Abstract

    9.1 Preparation of Axoplasm

    9.2 Analysis of Axoplasmic Microtubule Dynamics

    9.3 Biochemistry of Axoplasmic Microtubules

    9.4 Immunohistochemistry of Axoplasmic Microtubules

    References

    Chapter 10. Imaging GTP-Bound Tubulin: From Cellular to In Vitro Assembled Microtubules

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 11. Tubulin-Specific Chaperones: Components of a Molecular Machine That Assembles the α/β Heterodimer

    Abstract

    Introduction

    References

    Chapter 12. Heterotrimeric G Proteins and Microtubules

    Abstract

    Introduction

    Concluding Comments

    Acknowledgments

    References

    Chapter 13. Purification and Biophysical Analysis of Microtubule-Severing Enzymes In Vitro

    Abstract

    Introduction

    Discussion and Summary

    Acknowledgments

    References

    Chapter 14. Measurement of In Vitro Microtubule Polymerization by Turbidity and Fluorescence

    Abstract

    14.1 Background and Theory

    14.2 Materials and Equipment

    14.3 Methods

    References

    Chapter 15. Live-Cell Imaging of Microtubules and Microtubule-Associated Proteins in Arabidopsis thaliana

    Abstract

    Introduction

    Concluding Comments

    Acknowledgment

    References

    Chapter 16. Investigating Tubulin Posttranslational Modifications with Specific Antibodies

    Abstract

    Introduction

    Concluding Remarks

    Acknowledgments

    References

    Chapter 17. Purification and Assembly of Bacterial Tubulin BtubA/B and Constructs Bearing Eukaryotic Tubulin Sequences

    Abstract

    Introduction and Rationale

    Conclusions

    Acknowledgments

    References

    Chapter 18. Microtubule-Associated Proteins and Tubulin Interaction by Isothermal Titration Calorimetry

    Abstract

    Introduction

    Conclusion

    Acknowledgment

    References

    Chapter 19. Methods for Studying Microtubule Binding Site Interactions: Zampanolide as a Covalent Binding Agent

    Abstract

    Introduction

    Summary

    Acknowledgments

    References

    Chapter 20. Studying Kinetochore-Fiber Ultrastructure Using Correlative Light-Electron Microscopy

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 21. Fluorescence-Based Assays for Microtubule Architecture

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 22. Structure–Function Analysis of Yeast Tubulin

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 23. Using MTBindingSim as a Tool for Experimental Planning and Interpretation

    Abstract

    Introduction

    Conclusion

    References

    Chapter 24. Imaging Individual Spindle Microtubule Dynamics in Fission Yeast

    Abstract

    Introduction

    Conclusion

    Acknowledgments

    References

    Index

    Volumes in Series

Product details

  • No. of pages: 440
  • Language: English
  • Copyright: © Academic Press 2013
  • Published: August 20, 2013
  • Imprint: Academic Press
  • Hardcover ISBN: 9780124077577
  • eBook ISBN: 9780124078888

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