Reconstituting the cytoskeleton:
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Amsterdam [u.a.]
Elsevier, Acad. Press
2014
|
| Edition: | 1. ed. |
| Series: | Methods in enzymology
540 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XXIII, 484 S., [15] Bl. Ill., graph. Darst. |
| ISBN: | 9780123979247 |
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MARC
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| 245 | 1 | 0 | |a Reconstituting the cytoskeleton |c ed. by Ronald D. Vale |
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Record in the Search Index
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| adam_text | Titel: Reconstituting the cytoskeleton
Autor: Vale, Ronald D
Jahr: 2014
CONTENTS
Contributors xiii
Preface xix
Section I
Polymer Dynamics
1. Actin Filament Oynamics Using Microfluidics 3
Marie-France Carlier, Guillaume Romet-Lemonne,
and Antoine Jégou
1. Introduction 4
2. Key Technical Aspects of the Microfluidlc Method 5
3. Assets for the Study of Actln Dynamics 10
4. Perspectives 13
References 15
2. Bacterial Actin-Like Proteins: Purification and Characterization
of Self-Assembly Properties 19
Natalie A. Petek and R. Dyche Mullins
1. Introduction 20
2. Conclusion 33
Acknowledgments 33
References 33
3. Quantitative Analysis of Microtubule Self-assembly Kinetics
and Tip Structure 35
Louis S. Prahl, Brian T. Castle, Melissa K. Gardner,
and David J. Odde
1. Introduction 36
2. Tip-Tracking Algorithm Walkthrough 39
3. In Vivo Tip-Tracking Protocol 41
4. TipTracker Optimization Using Fixed Cells 47
5. Considerations for In Vitro Assays and Other Filament Systems 47
Acknowledgments 50
References 50
vi
Contents
Section II
Polymer Nudeation and Regulation
4. Biochemical Reconstitution of the WAVE Regulatory Complex 55
Baoyu Chen, Shae B. Padrick, Lisa Henry, and Michael K. Rosen
1. Introduction 56
2. Overview of the In Vitro Reconstitution Method 59
3. Key Reagents 61
4. Reconstitution of the WAVE/Abi/HSPC300 Trimer 61
5. Reconstitution of the Sra 1 /Napl Dimer 65
6. Reconstitution of the WRC Pentamer 68
Acknowledgments 70
References 70
5. Rotational Movement of Formins Evaluated by Using
Single-Molecule Fluorescence Polarization 73
Hiroaki Mizuno and Naoki Watanabe
1. Introduction 74
2. Theoretical Concepts and Optics for Single-Molecule
Fluorescence Polarization 75
3. Proteins 80
4. Observation of Rotational Movement of TMR-F-Actin 84
5. Limiting Axial Rotation of F-Actin Using Biotinylated Actin 88
6. Data Analysis 92
7. Perspectives 93
Acknowledgments 93
References 93
6. Single-Molecule Studies of Actin Assembly and
Disassembly Factors 95
Benjamin A. Smith, Jeff Gelles, and Bruce L. Goode
1. Introduction 96
2. Protein Tags, Labeling, and Tethering 98
3. Labeling and Surface Confinement of Actin Filaments 99
4. Slide Surface Treatment 100
5. Requirements for Single-Molecule Detection 101
6. Microscope Considerations 101
7. Imaging Actin Filaments and Single Molecules 103
Contents
v¡¡
8. Dual-Color TIRF Imaging of Actln-Regulatory Mechanisms 106
9. Measuring Kinetics of Single-Molecule Interactions with Actin Filaments 108
10. Coordination of Multiple Actin-Regulatory Factors from
Multicolor Imaging 112
Acknowledgments 114
References 114
7. Assaying Microtubule Nucleation by the 7-Tubulin
Ring Complex 119
Yuk-Kwan Choi and Robert Z. Qi
1. Introduction 120
2. yTuRC Isolation 121
3. Electron Microscopy 124
4. Microtubule Nucleation 125
5. Summary 126
Acknowledgments 128
References 128
8. Reconstituting Dynamic Microtubule Polymerization
Regulation by TOG Domain Proteins 131
Jawdat Al-Bassam
1. Introduction 132
2. Methods 134
3. Conclusion 146
Acknowledgments 147
References 147
9. Generation of Differentially Modified Microtubules
Using In Vitro Enzymatic Approaches 149
Annapurna Vemu, Christopher P. Garnham, Duck-Yeon Lee,
and Antonina Roll-Mecak
1. Introduction 150
2. Purification and Characterization of Unmodified Microtubules 153
3. Purification of Tubulin Modification Enzymes: a-TAT, TTLL7, and TTL 154
4. Generation of Differentially Modified Tubulin and Microtubules 157
5. Conclusions 163
References 164
viii
Contents
Section III
Molecular Motor Ensembles on Natural
and Engineered Cargoes
10. Engineering Defined Motor Ensembles with DNA Origami 169
Brian S. Goodman and Samara L. Reek-Peterson
1. Introduction 170
2. Building the DNA Origami Structure 171
3. Covalent Attachment of Oligonucleotides to Motor Proteins 176
4. Forming the Motor-DNA Origami Complex 181
5. Single-Molecule Motility Analysis of Motor-DNA Origami
Complexes 182
6. Data Analysis Considerations 185
7. Summary and Future Directions 185
Acknowledgments 186
References 187
11. Construction and Analyses of Elastically Coupled
Multiple-Motor Systems 189
Arthur Rogers, Pamela E. Constantinou, D. Kenneth Jamison,
Jonathan W. Driver, and Michael R. Diehl
1. Introduction 190
2. Synthetic Strategy: A Modular Plug-and-Play Approach 191
3. Two-Motor Biophysical Assays 197
4. Concluding Remarks and Future Perspectives 202
Acknowledgments 202
References 203
12. Reconstitution of Cortical Dynein Function 205
Sophie Roth, Liedewij Laan, and Marileen Dogterom
1. Introduction 206
2. Reconstitution of Cortical Dynein Function in 1D Geometries 207
3. Reconstitution of Cortical Dynein Function in 2D Geometries 215
4. Reconstitution of Cortical Dynein Function In 3D Geometries 219
5. Discussion and Perspectives 226
Acknowledgments 228
References 228
Contents ¡X
13. Reconstitution of Microtubule-Dependent Organelle
Transport 231
Pradeep Barak, Ashim Rai, Alok Kumar Dubey, Priyanka Rai,
and Roop Mallik
1. Introduction 232
2. Isolation and Motility of LBPs from Dictyostelium 234
3. Isolation and Motility of LDs from Rat Liver 238
4. Conclusions 245
Acknowledgments 246
References 246
14. Reconstituting the Motility of Isolated Intracellular
Cargoes 249
Adam G. Hendricks, Yale E. Goldman, and Erika L. F. Holzbaur
1. introduction 250
2. Isolation of Neuronal Transport Vesicles 250
3. Isolation of Latex Bead Compartments 253
4. In Vitro Motility Assays 256
5. Imaging and Analysis 257
6. Troubleshooting 259
7. Comparison to Measurements in Living Cells 259
8. Outlook 260
Acknowledgments 260
References 260
Section IV
Building Higher Order Networks and Interactions
15. Reconstitution of Contractile Actomyosin Arrays
Michael Murrell, Todd Thoresen, and Margaret Gardel
1. Introduction
2. Reagents
3. Reconstituted Actomyosin Bundles
4. Biomimetic Cortex
5. Future Directions
Acknowledgment
References
265
266
267
268
272
278
281
281
X
Contents
16. Directed Actin Assembly and Motility 283
Rajaa Boujemaa-Paterski, Rémi Galland, Cristian Suarez, Christophe Guérin,
Manuel Théry, and Laurent Blanchoin
1. Introduction 284
2. Reconstitution of Actin-Based Motility in a G-Actin-Buffered
Medium 285
3. Assembly of Branched Actin Networks on a Wide Range of Geometries 289
4. Reconstitution of 3D Connections of Structured Actin Networks 294
5. Concluding remarks 298
Acknowledgments 298
References 298
17. In Vitro Reconstitution of Dynamic Microtubules Interacting
with Actin Filament Networks 301
Magdalena Preciado López, Florian Huber, llya Grigoriev, Michel 0.
Steinmetz, Anna Akhmanova, Marileen Dogterom, and Gijsje H. Koenderink
1. Introduction 302
2. Experimental Methods 305
3. Outlook 316
4. Reagents and Stocks 316
Acknowledgments 317
References 318
18. Measuring Kinetochore-Microtubule Interaction In Vitro 321
Jonathan W. Driver, Andrew F. Powers, Krishna K. Sarangapani,
Sue Biggins, and Charles L. Asbury
1. Introduction 322
2. The Leap to In Vitro: Purification of Intact, Functional Kinetochores 323
3. The Assays 327
4. Summary 335
Acknowledgments 336
References 336
19. Micropattern-Guided Assembly of Overlapping Pairs of
Dynamic Microtubules 339
Franck J. Fourniol, Tai-De Li, Peter Bieling, R. Dyche Mullins,
Daniel A. Fletcher, and Thomas Surrey
1. Introduction 340
2. Reagents and Equipment 341
Contents
xi
3. Glass Treatment and Surface Chemistry 346
4. Microtubule Overlap Assay on Micropattern 353
5. Discussion 358
Acknowledgments 359
References 359
Section V
Cell Extract Systems
20. WAVE Regulatory Complex Activation 363
Peter J. Hume, Daniel Humphreys, and Vassilis Koronakis
1. Introduction 364
2. Generating Phospholipid Bilayer-Coated Silica Microspheres 365
3. Preparation of Brain Extract 368
4. Assay of WAVE-Dependent Motility by Phospholipid-Coated Microspheres 370
5. Reconstituting Activation of WRC by Cooperating Arf and Rac1 GTPases 372
6. Concluding Remarks 377
Acknowledgments 378
References 378
21. Dissecting Principles Governing Actin Assembly Using
Yeast Extracts 381
Alphée Michelot and David G. Drubin
1. Introduction 382
2. Yeast Culture and Extract Preparation 383
3. Surface Protein Activation 388
4. Actin Assembly from Yeast Extracts 389
5. Deducing Protein Functions with Mutant Strains 393
6. Perspectives 395
Acknowledgments 395
References 396
22. Xenopus Egg Cytoplasm with Intact Actin 399
Christine M. Field, Phuong A. Nguyen, Keisuke Ishihara,
Aaron C. Groen, and Timothy J. Mitchison
1. Introduction 400
2. Actin-lntact Extract Preparation 401
3. Cell Cycle-Regulated Actomyosln Contractility 411
4. Cell-Free Reconstitution of l-Phase Aster Growth and Interaction 412
Acknowledgments 413
References 414
xii
Contents
23. Glycogen-Supplemented Mitotic Cytosol for Analyzing Xenopus
Egg Microtubule Organization 417
Aaron C. Groen, Phuong A. Ngyuen, Christine M. Field, Keisuke Ishihara,
and Timothy J. Mitchison
1. Introduction 418
2. Development of the Cytosol System 420
3. Preparation of Glycogen-Supplemented Cytosol 421
4. Assay of Ran-Promoted Microtubule Assembly 423
5. Self-Organization of Spindle-Like Microtubule Arrays 424
6. Antibody-Based Fluorescent Probes 428
7. Future Directions 430
Acknowledgments 432
References 432
24. Spindle Assembly on Immobilized Chromatin Micropatterns 435
Céline Pugieux, Serge Dmitrieff, Katarzyna Tarnawska, and François Nédélec
1. Introduction 436
2. Overview of the Method 437
3. Preparative Steps 437
4. Patterning 440
5. Spindle Arrays 442
6. Analysis 443
7. Conclusions 447
Acknowledgments 443
References 443
Author Index
Subject Index
449
471
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| spelling | Reconstituting the cytoskeleton ed. by Ronald D. Vale 1. ed. Amsterdam [u.a.] Elsevier, Acad. Press 2014 XXIII, 484 S., [15] Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Methods in enzymology 540 Zellskelett (DE-588)4121944-2 gnd rswk-swf Rekonstitution (DE-588)4177651-3 gnd rswk-swf 1\p (DE-588)4143413-4 Aufsatzsammlung gnd-content Zellskelett (DE-588)4121944-2 s Rekonstitution (DE-588)4177651-3 s DE-604 Vale, Ronald D. Sonstige oth Methods in enzymology 540 (DE-604)BV000000938 540 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027337943&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Reconstituting the cytoskeleton Methods in enzymology Zellskelett (DE-588)4121944-2 gnd Rekonstitution (DE-588)4177651-3 gnd |
| subject_GND | (DE-588)4121944-2 (DE-588)4177651-3 (DE-588)4143413-4 |
| title | Reconstituting the cytoskeleton |
| title_auth | Reconstituting the cytoskeleton |
| title_exact_search | Reconstituting the cytoskeleton |
| title_full | Reconstituting the cytoskeleton ed. by Ronald D. Vale |
| title_fullStr | Reconstituting the cytoskeleton ed. by Ronald D. Vale |
| title_full_unstemmed | Reconstituting the cytoskeleton ed. by Ronald D. Vale |
| title_short | Reconstituting the cytoskeleton |
| title_sort | reconstituting the cytoskeleton |
| topic | Zellskelett (DE-588)4121944-2 gnd Rekonstitution (DE-588)4177651-3 gnd |
| topic_facet | Zellskelett Rekonstitution Aufsatzsammlung |
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