Main description:

This volume provides state-of-the-art methodology to functionalize microtubules and construct microtubule-based materials. Chapters consists of six sections detailing modification of microtubule structures, observation and control of microtubule movement, material applications of microtubules in vitro, development of microtubule-binding molecules, comprehensive approaches to analyze properties of microtubules, and functionalization of microtubules in living cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Microtubules: Methods and Protocols aims to provide useful information for construction of microtubule-based materials and help to create new ideas for the next-generation of materials and applications through detailed protocols.

Contents:

PART I: Preparation and Modification of Microtubules

1. Purification of Tubulin from Porcine Brain and its Fluorescence Dye Modification

Satsuki Ishii, Mousumi Akter, Jakia Jannat Keya, Mst. Rubaya Rashid, Farhana Afroze, Syeda Rubaiya Nasrin, and Akira Kakugo

2. Facile Method of Tubulin Purification from Goat Brain for Reconstitution of Microtubule Associated Intracellular Function

Satyajit Ghosh, Shubham Garg, Nabanita Mukherjee, and Surajit Ghosh

3. Functionalization of Tubulin: Approaches to Modify Tubulin with Biotin and DNA

Mousumi Akter, Jakia Jannat Keya, Arif Md. Rashedul Kabir, Mst. Rubaya Rashid, Satsuki Ishii, and Akira Kakugo

4. Electro-Modulation of Tubulin Properties and Function

Djamel Eddine Chafai and Michal Cifra

PART II: Observation and Control of Microtubule Movement

5. In Vitro Reconstitution of Microtubule Dynamics and Severing Imaged by Label-Free Interference Reflection Microscopy

Yin-wei Kuo and Jonathon Howard

6. Characterizing the Number of Kinesin Motors Bound to Microtubules in the Gliding Motility Assay using FLIC Microscopy

Virginia VanDelinder and George D. Bachand

7. Design of Mechanical and Electrical Properties for Multi-Directional Control of Microtubules

Hang Zhou, Taikopaul Kaneko, Naoto Isozaki, and Ryuji Yokokawa

8. Linear-Zero Mode Waveguides for Single-Molecule Fluorescence Observation of Nucleotides in Kinesin-Microtubule Motility Assay

Kazuya Fujimoto, Ryota Iino, and Ryuji Yokokawa

9. Microtubules and Quantum Dots Integration Leads to Conjugates with Applications in Biosensors and Bionanodevices

Xiao Hu and Cerasela Zoica Dinu

PART III: Microtubule-Based Active Matters

10. Assembling Microtubule-Based Active Matter

Alexandra M. Tayar, Linnea M. Lemma, and Zvonimir Dogic

11. Spontaneous Alignment of Microtubules via Tubulin Polymerization in a Narrow Space under a Temperature Gradient

Kazuhiro Shikinaka

12. Dynamic Pattern Formation of Active Matters Triggered by Mechanical Stimuli

Jakia Jannat Keya, Arif Md. Rashedul Kabir, Mousumi Akter, and Akira Kakugo

13. Spontaneously Beating Biomimetic Structures

Isabella Guido

14. Construction of Molecular Robots from Microtubules for Programmable Swarming

Jakia Jannat Keya, Mousumi Akter, Arif Md. Rashedul Kabir, Mst. Rubaya Rashid, and Akira Kakugo

15. Fabrication of Artificial Muscle from Microtubules, Kinesins and DNA Origami Nanostructures

Jakia Jannat Keya, Mousumi Akter, Arif Md. Rashedul Kabir, Satsuki Ishi, and Akira Kakugo

PART IV: Microtubule-Binding Molecules

16. Encapsulation of Nanomaterials Inside Microtubules by Using a Tau-derived Peptide

Hiroshi Inaba and Kazunori Matsuura

17. Investigating Tubulin-Drug Interaction Using Fluorescence Spectroscopy

Anuradha Kumari and Dulal Panda

PART V: Analysis of Microtubule Dynamics and Structures

18. Visualization and Quantification of Microtubule Self-repair

Jeremie Gaillard, Laurent Blanchoin, Manuel Thery, and Laura Schaedel

19. Cargo Transport by Microtubule-Associated Motor Protein Along Mechanically Deformed Microtubules

Syeda Rubaiya Nasrin, Arif Md. Rashedul Kabir, and Akira Kakugo

20. Mechanical Deformation of Microtubules on a Two-Dimensional Elastic Medium

Syeda Rubaiya Nasrin, Farhana Afroze, Arif Md. Rashedul Kabir, and Akira Kakugo

21. Real-Time Imaging of Single TuRC-Mediated Microtubule Nucleation Events In Vitro by TIRF Microscopy

Tanja Consolati, Gil Henkin, Johanna Roostalu, and Thomas Surrey

22. Microtubule Preparation for Investigation with High-Speed Atomic Force Microscopy

Christian Ganser and Takayuki Uchihashi

23. Crystallization Systems for the High-Resolution Structural Analysis of Tubulin-Ligand Complexes

Tobias Muhlethaler, Natacha Olieric, Valentin A. Ehrhard, Maximilian Wranik, Joerg Standfuss, Ashwani Sharma, Andrea E. Prota, and Michel O. Steinmetz

24. Cryo-EM Visualization of Neuronal Particles Inside Microtubules

Sylvie Gory-Faure, Julie Delaroche, Camille Cuveillier, Christian Delphin, and Isabelle Arnal

25. Reconstituting the Interaction Between Purified Nuclei and Microtubule Network

Gokce Agsu, Jeremie Gaillard, Bruno Cadot, Laurent Blanchoin, Emmanuelle Fabre, and Manuel Thery

PART VI: Modulation of Cellular Microtubules

26. Photocontrolling Microtubule Dynamics with Photoswitchable Chemical Reagents

Oliver Thorn-Seshold and Joyce C.M. Meiring

27. Monitoring the Disruptive Effects of Tubulin Binding Agents on Cellular Microtubules

Anuradha Kumari and Dulal Panda

28. Pacific Blue-Taxoids as Fluorescent Molecular Probes of Microtubules

Angelo E. Andres, Digamber Rane, and Blake R. Peterson

29. Controlling Cell Shape and Microtubule Organization by Extracellular Matrix Micropatterning

Alessandro Dema, Shima Rahgozar, Laurent Siquier, Jeffrey van Haren, and Torsten Wittmann