Regulators and Effectors of Small GTPases, Part D: Rho Family (eBook)

Cover 1
Table of Contents 6
Contributors to Volume 325 10
Preface 16
Volumes in Series 18
Section I: Purification, Posttranslational Modification, and in Vitro Regulation 37
Chapter 1. Purification and Biochemical Characterization of TC10 39
Chapter 2. Expression and Purification of Rho/RhoGDI Com- plexes 51
Chapter 3. Bacterial Expressed DH and DH/PH Domains 61
Chapter 4. Biochemical Analysis of Regulation of Vav, a Guanine-Nucleotide Exchange Factor for Rho Family of GTPases 74
Chapter 5. Activation of Rac1 by Human Tiam1 87
Chapter 6. Activation of Rho GEF Activity by Ga13 97
Chapter 7. Purification and Characterization of Guanine Nucleotide Dissociation Stimulator Protein 107
Chapter 8. Purification and Biochemical Activity of Salmonella Exchange Factor SopE 118
Chapter 9. Stimulation of Rho GDI Release by ERM Proteins 127
Chapter 10. Expression and Activity of Human Prenylcysteine- Directed Carboxyl Methyltransferase 137
Chapter 11. Purification and Evaluation of Large Clostridial Cytotoxins That Inhibit Small GTPases of Rho and Ras Subfamilies 150
Chapter 12. Rho GTPase-Activating Toxins: Cytotoxic Necrotizing Factors and Dermonecrotic Toxin 161
Section II: Purification and Activity of GTPase Targets 173
Chapter 13. Measurement of GTPase · Effector Affinities 175
Chapter 14. Purification and in Vitro Activity of Rho-Associated Kinase 185
Chapter 15. Purification and in Vitro Activities of p21-Activated Kinases 191
Chapter 16. Stimulation of Phospholipase C-ß2 by Rho GTPases 203
Chapter 17. Regulation of Phospholipase D1 Activity by Rho GTPases 213
Chapter 18. In Vitro Interaction of Phosphoinositide-4-phosphate 5-Kinases with Rac 226
Chapter 19. Activity and Regulation of p35/Cdk5 Kinase Complex 236
Chapter 20. Actin Assembly Mediated by Arp2/3 Complex and WASP Family Proteins 250
Chapter 21. In Vitro Actin Polymerization Using Polymorphonuclear Leukocyte Extracts 273
Section III: Analysis of Rho GTPase Function 291
Chapter 22. Determination of GTP Loading on Rac and Cdc42 in Platelets and Fibroblasts 293
Chapter 23. Determination of GTP Loading on Rho 300
Chapter 24. Use and Properties of ROCK-Specific Inhibitor Y-27632 309
Chapter 25. Inducible Membrane Recruitment of Small GTP-Binding Proteins by Rapamycin-Based System in Living Cells 321
Chapter 26. Expression of Rho GTPases Using Retroviral Vectors 331
Chapter 27. Expression of Rho GTPases Using Adenovirus Vectors 339
Chapter 28. In Vivo Activity of Wild-Type and Mutant PAKs 351
Chapter 29. Single Cell Assays for Rac Activity 363
Chapter 30. Effect of Rho GTPases on Na–H Exchanger in Mammalian Cells 370
Chapter 31. Actin Filament Assembly in Permeabilized Platelets 385
Chapter 32. Rho GTPases: Secretion and Actin Dynamics in Permeabilized Mast Cells 392
Chapter 33. Isolation and in Vitro Contraction of Stress Fibers 405
Chapter 34. Inhibition of Rho GTPases Using Protein Geranylgeranyltransferase I Inhibitors 417
Chapter 35. Imaging Spatiotemporal Dynamics of Rac Activation in Vivo with FLAIR 425
Section IV: Biological Assays of Rho GTPase Function 437
Chapter 36. Rho-like GTPases in Tumor Cell Invasion 439
Chapter 37. Mouse Embryo Fibroblasts. A Genetic Model System for Studying Rho- and Ras-Dependent Cell Cycle Progression 451
Chapter 38. Analyses of Transforming Activity of Rho Family Activators 461
Chapter 39. Rho GTPases and Cell Migration–Fibroblast Wound Healing 477
Chapter 40. Rho GTPases and Cell Migration: Measurement of Macrophage Chemotaxis 485
Chapter 41. Rho GTPases and Macrophage Phagocytosis 498
Chapter 42. Rho GTPases and Axonal Growth Cone Collapse 509
Chapter 43. Study of in Situ Function of Cytoskeletal Proteins in Lamellipodia and Filopodia Using Microscale Chromophore-Assisted Laser Inactivation 518
Chapter 44. Modulation of Actin Cytoskeleton by Salmonella GTPase Activating Protein SptP 532
Author Index 541
Subject Index 567