Table of contents : Table of Contents......Page 6 Contributors to Volume 256......Page 10 Preface......Page 13 Section I Expression. Purification. and Posttranslational Modification......Page 14 [ 1] Purification of Recombinant Rho / Rac / G25K from Escherichia coli......Page 16 [2] Purification of Baculovirus-Expressed Cdc42Hs......Page 24 [3] Purification and Properties of Rac2 from Human Leukemia Cells......Page 28 [4] Purification of Rac2 from Human Neutrophils......Page 38 [5] Purification of Rac-GDP Dissociation Inhibitor Complex from Phagocyte Cytosol......Page 46 [6] Purification and Properties of Recombinant Rho-GDP Dissociation Inhibitor......Page 54 [7] Prenylcysteine-Directed Carboxyl Methyltransferase Activity in Human Neutrophil Membranes......Page 62 Section II Guanine Nucleotide Exchange and Hydrolysis......Page 78 [8] Measurement of Intrinsic Nucleotide Exchange and GTP Hydrolysis Rates......Page 80 [9] Guanine Nucleotide Exchange Catalyzed by dbl Oncogene Product......Page 90 [10] Stimulation of Nucleotide Exchange on Ras- andRho-Related Proteins by Small GTP-Binding Protein GDP Dissociation Stimulator......Page 98 [11] Interaction of Ect2 and Dbl with Rho-Related GTPases......Page 103 [13] Purification and GTPase-Activating Protein Activity of Baculovirus Expressed p 190......Page 118 [ 14] GTPase-Activating Protein Activityof n(al)-Chimaerin and Effect of Lipids......Page 127 [15] Characterization of Breakpoint Cluster Region Kinase and SH2-Binding Activities......Page 138 [16] Identification of GTPase-Activating Proteins by Nitrocellulose Overlay Assay......Page 143 [17] Identification of 3BP- 1 in cDNA Expression Libraryby SH3 Domain Screening......Page 153 Section III Cell Expression and in Vitro Analysis......Page 162 [18] Serum Induction of RhoG Expression......Page 164 [19] Microinjection of Epitope-Tagged Rho Family cDNAs and Analysis by Immunolabeling......Page 175 [20] Purification and Assay of Recombinant C3 Transferase......Page 187 [21] In Vitro ADP-Ribosylation of Rho by Bacterial ADP-Ribosyltransferases......Page 197 [22] Preparation of Native and Recombinant Clostridiumbotulinum C3 ADP-Ribosyltransferase and Identification of Rho Proteins by ADP-Ribosylation......Page 209 [23] In Vitro Binding Assay for Interactions of Rho and Rac with GTPase-Activating Proteins and Effectors......Page 220 [24] Purification and Assay of Kinases That Interact with Rac / Cdc42......Page 228 [25] Yeast Two-Hybrid System to Detect Protein-Protein Interactions with Rho GTPases......Page 241 [26] Assay for Rho-Dependent Phosphoinositide 3-Kinase Activity in Platelet Cytosol......Page 254 [27] Neutrophil Phospholipase D: Inhibition by Rho-GDP Dissociation Inhibitor and Stimulation by Small GTPase GDP Dissociation Stimulator......Page 259 [28] Measurement of Rac Translocation from Cytosol to Membranes in Activated Neutrophils......Page 269 [29] Reconstitution of Cell-Free NADPH Oxidase Activity by Purified Components......Page 281 Section IV. Biological Activity......Page 292 [30] Genetic and Biochemical Analysis of Cdc42p Function in Saccharomyces cerevisiae and Schizosaccharomyces pombe......Page 294 [31] Lymphocyte Aggregation Assay and Inhibition by Clostridium botulinum C3 ADP-Ribosyltransferase......Page 303 [32] Inhibition of p21 Rho in Intact Cells by C3 Diphtheria Toxin Chimera Proteins......Page 310 [33] Growth Factor-Induced Actin Reorganization in Swiss 3T3 Cells......Page 319 [34] Microinjection of Rho and Rac into Quiescent Swiss 3T3 Cells......Page 326 [35] Inhibition of Lymphocyte-Mediated Cytotoxicity by Clostridium botulinum C3 Transferase......Page 333 [36] Neutrophil Chemotaxis Assay and Inhibition by C3 ADP-Ribosyltransferase......Page 340 [37] Cell Motility Assay and Inhibition by Rho-GDP Dissociation Inhibitor......Page 349 [38] Cell Transformation by dbl Oncogene......Page 360 [39] Inhibition of Rac Function Using Antisense Oligonucleotides......Page 371 Author Index......Page 380 Subject Index......Page 398