Methods in Enzymology (Hardcover)

siRNA cell arrays for high-content screening microscopy; Imaging platforms for measurement of membrane trafficking; Use of the harvestorR data base to annotate the human proteome for GTPase function; Microinjection as a tool to explore Small GTPase function; Visualizing intracellular events in vivo by combined video fluorescence and 3-D electron microscopy; Recombinant production in Baculovirus-infected insect cells and purification of the mammalian Sec13/Sec31 complex; Purification and properties of mammalian Sec23/24 from insect cells; Purification and functional properties of yeast Sec12 GEF; Reconstitution of cargo-dependent COPII coat assembly on proteoliposomes; Assay of COPII vesicle formation using a real-time fluorescence assay; Assay and measurement of phospholipase D activation by Sar1; Use of Acyltransferase Inhibitors to Block Vesicular Traffic Between the ER and Golgi Complex; Tethering assays for COPI Vesicles Mediated by Golgins; Investigating the Role of ADP-Ribosylation Factor 6 in Tumor Cell Invasion and Extracellular Signal-Regulated Kinase Activation; Assay and properties of the Arf GAPs AGAP1, ASAP1 and ArfGAP1; Preparation of myristoylated ARF1 and ARF6; BIG1 and BIG2, Brefeldin A-inhibited Guanine Nucleotide-exchange Factors for ADP-Ribosylation Factors (ARFs); Cytohesin-1: Structure, function, and ARF activation; ADP-Ribosylation Factor Domain Protein 1 (ARD1), a Multifunctional Protein with Ubiquitin E3 Ligase, GAP, and ARF Domains; Expression of BIG2 and Analysis of its Function in Mammalian Cells; Assay and properties of the ARF-GAP, AMAP2, in ARF6 function; Functional assay of EFA6A, a Guanine Nucleotide Exchange Factor for ADP-ribosylation Factor 6 (ARF6), in Dendritic Formation of Hippocampal Neurons; Functional assay of ARNO and ARF6 in neurite elongation and branching; Assay and functional properties of IPCEF1 as regulator of cytohesin GEFs; Assay and properties of the GIT1/p95-APP1 ARFGAP; Preparation and Characterization of Recombinant Golgin Tethers; Purification and functional properties of CtBP/Bars in membrane fission; In vitro Assays of Arf1 interaction with GGA proteins; The role of EFA6, Exhange Factor for Arf6, for tight junction assembly, functions and interaction with actin cytoskeleton; In vitro reconstitution of ARF-regulated cytoskeletal dynamics on Golgi membranes; Assay and properties of Arfaptin 2 binding to Rac1 and ADP-ribosylation Factors (ARFs); Analysis of ARF Interaction with GGAs In Vitro and In Vivo; Arf6 modulates the beta actin-specific regulator, betacap73; Functional assay of effectors of ADP ribosylation factor 6 during clathrin/AP-2 coat recruited of membranes; Assays to study Phospholipase D regulation by inositol phospholipids and ADP-ribosylation factor 6; Assay and functional properties of the tyrosine kinase Pyk2 in regulation of ARF1 through ASAP1 phosphorylation; ARF6 regulation of PI(4,5)P2 synthesis, endocytosis and exocytosis; Interaction of Arl1 GTPase with the GRIP domain of Golgin-245 as assessed by GST (glutathione-S-transferase) pull-down experiments; Functional analysis of Arl1 and Golgin-97 in endosome-to-TGN transport using recombinant Shiga toxin B fragment; Assays used in the analysis of ARL2 and its binding partners;
Assay and functional analysis of the ARL3 effector RP2 involved in X-linked retinitis pigmentosa; Reconstitution of transport to recycling endosomes in vitro; Robust colorimetric assays for dynamin's basal and stimulated GTPase activities; Clathrin-coated vesicle formation from isolated plasma membranes; Assay of biochemical properties of human guanylate-binding protein 1; Stimulation of Dynamin GTPase activity by amphiphysin; Purification and properties of Tuba, a novel dynamin-actin effector; Expression and properties of sorting nexin 9 in dynamin-mediated endocytosis; Rapid purification of native dynamin I and colorimetric GTPase assay;
Assay and functional properties of auxilin-dynamin interactions; Assay and functional analysis DLP1 in peroxisome division; In vitro reconstitution of discrete stages of dynamin-dependent endocytosis; A continuous regenerative coupled GTPase assay for dynamin-related problems; Assay and properties of the mitochondrial dynamin related protein Opa1; Assay and functional analysis of dynamin-like Mx proteins