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

Cover 1
Copyright Page 5
Table of Contents 6
Contributors to Volume 406 12
Preface 20
Volumes in Series 22
Chapter 1: Purification and Biochemical Properties of Rac1, 2, 3 and the Splice Variant Rac1b 45
Chapter 2: Biochemical Analyses of the Wrch Atypical Rho Family GTPases 55
Chapter 3: Purification of P-Rex1 from Neutrophils and Nucleotide Exchange Assay 70
Chapter 4: In Vitro Guanine Nucleotide Exchange Activity of DHR-2/DOCKER/CZH2 Domains 85
Chapter 5: Biochemical Characterization of the Cool (Cloned-Out-of-Library)/Pix (Pak-Interactive Exchange Factor) Proteins 102
Chapter 6: GEF and Glucosylation Assays on Liposome-Bound Rac 114
Chapter 7: Phosphorylation of RhoGDI by p21-Activated Kinase 1 124
Chapter 8: Purification of ARAP3 and Characterization of GAP Activities 135
Chapter 9: Regulation of RhoGAP Specificity by Phospholipids and Prenylation 148
Chapter 10: Purification and Activity of the Rho ADP-Ribosylating Binary C2/C3 Toxin 161
Chapter 11: Purification of TAT-C3 Exoenzyme 172
Chapter 12: Imaging and Photobleach Correction of Mero-CBD, Sensor of Endogenous Cdc42 Activation 184
Chapter 13: Cdc42 and PI(4,5)P2-Induced Actin Assembly in Xenopus Egg Extracts 200
Chapter 14: In Vitro Reconstitution of Cdc42-Mediated Actin Assembly Using Purified Components 218
Chapter 15: Biochemical Analysis of Mammalian Formin Effects on Actin Dynamics 234
Chapter 16: Formin Proteins: Purification and Measurement of Effects on Actin Assembly 259
Chapter 17: Purification and Kinase Assay of PKN 278
Chapter 18: Purification and Enzyme Activity of ACK1 294
Chapter 19: Direct Activation of Purified Phospholipase C Epsilon by RhoA Studied in Reconstituted Phospholipid Vesicles 304
Chapter 20: Regulation of PLCbeta Isoforms by Rac 316
Chapter 21: Biochemical Properties and Inhibitors of (N-)WASP 325
Chapter 22: The Use of GFP to Localize Rho GTPases in Living Cells 340
Chapter 23: Analysis of the Spatiotemporal Activation of Rho GTPases Using Raichu Probes 359
Chapter 24: Measurement of Activity of Rho GTPases During Mitosis 376
Chapter 25: Inhibition of Rho GTPases by RNA Interference 389
Chapter 26: RNA Interference Techniques to Study Epithelial.Cell Adhesion and Polarity 406
Chapter 27: Nucleofection of Primary Neurons 418
Chapter 28: Dock180-ELMO Cooperation in Rac Activation 432
Chapter 29: Rho GTPase Activation by Cell-Cell Adhesion 446
Chapter 30: Activation of Rap1, Cdc42, and Rac by Nectin Adhesion System 459
Chapter 31: Analysis of Activated GAPs and GEFs in Cell Lysates 469
Chapter 32: Degradation of RhoA by Smurf1 Ubiquitin Ligase 481
Chapter 33: Ubiquitin-Mediated Proteasomal Degradation of Rho Proteins by the CNF1 Toxin 491
Chapter 34: Regulation of Superoxide-Producing NADPH Oxidases in Nonphagocytic Cells 500
Chapter 35: Activation of MEKK1 by Rho GTPases 512
Chapter 36: Activation of the Apoptotic JNK Pathway Through the Rac1-Binding Scaffold Protein POSH 523
Chapter 37: Quantification of Isozyme-Specific Activation of Phospholipase C-beta2 by Rac GTPases and Phospholipase C-epsiv by Rho GTPases in an Intact Cell Assay System 533
Chapter 38: Activation of Rho and Rac by Wnt/Frizzled Signaling 544
Chapter 39: Fluorescent Assay of Cell-Permeable C3 Transferase Activity 556
Chapter 40: Use of TIRF Microscopy to Visualize Actin and Microtubules in Migrating Cells 564
Chapter 41: Inhibition of ROCK by RhoE 577
Chapter 42: Conditional Regulation of a ROCK-Estrogen Receptor Fusion Protein 585
Chapter 43: Rational Design and Applications of a Rac GTPase-Specific Small Molecule Inhibitor 598
Chapter 44: In Vitro Assay of Primary Astrocyte Migration as a Tool to Study Rho GTPase Function in Cell Polarization 609
Chapter 45: Real-Time Centrosome Reorientation During Fibroblast Migration 623
Chapter 46: Lentiviral Delivery of RNAi in Hippocampal Neurons 637
Chapter 47: Methods for Studying Neutrophil Chemotaxis 649
Chapter 48: Measurement of Epidermal Growth Factor Receptor Turnover and Effects of Cdc42 658
Chapter 49: Tumor Cell Migration in Three Dimensions 669
Chapter 50: An In Vitro Model to Study the Role of Endothelial Rho GTPases During Leukocyte Transendothelial Migration 687
Chapter 51: Analysis of a Mitotic Role of Cdc42 700
Chapter 52: Plexin-Induced Collapse Assay in COS Cells 709
Chapter 53: Morphological and Biochemical Analysis of Rac1 in Three-Dimensional Epithelial Cell Cultures 720
Chapter 54: Using Three-Dimensional Acinar Structures for Molecular and Cell Biological Assays 736
Chapter 55: TC10 and Insulin-Stimulated Glucose Transport 745
Chapter 56: GTPases and the Control of Neuronal Polarity 759
Chapter 57: In Vitro Assembly of Filopodia-Like Bundles 771
Author Index 785
Subject Index 821