Table of Contents

Section I. Rab GTPases

1: Structural Basis for Rab Function: An Overview

2: Expression of Wild-Type and Mutant Green Fluorescent Protein-Rabl for Fluorescence Microscopy Analysis

3: Fluorescence Methods for Monitoring Interactions of Rab Proteins with Nucleotides, Rab Escort Protein, and Geranylgeranyltransferase

4: Prenylation of Rab Proteins in Vitro by Geranylgeranyltransferases

5: Antibody and Oligonucleotide Probes to Distinguish Intracellular Expression and Localization Patterns of Rab GDP-Dissociation Inhibitor Isoforms

6: Expression, Purification, and Biochemical Properties of Ypt/Rab GTPase-Activating Proteins of Gyp Family

7: Purification and Properties of Rab3 GDP/GTP Exchange Protein

8: Purification and Properties of Rab3 GTPase-Activating Protein

9: Rabphilin-3: A Target Molecule for Rab3 Small G Proteins

10: Doc2α as Modulator of Ca2+-Dependent Exocytosis

11: Purification and Properties of a GTPase-Activating Protein for Yeast Rab GTPases

12: Purification and Characterization of Yeast Exocyst Complex

13: Expression and Properties of Rab4 and Its Effector Rabaptin-4 in Endocytic Recycling

14: Purification of EEA1 from Bovine Brain Cytosol Using Rab5 Affinity Chromatography and Activity Assays

15: Expression, Purification, and Characterization of Rab5 Effector Complex, Rabaptin-5/Rabex-5

16: Measurement of Rab5 Protein Kinase B/akt and Regulation of Ras-Activated Endocytosis

17: Expression, Purification, and Biochemical Properties of Rabkinesin-6 Domains and Their Interactions with Rab6A

18: Expression and Functional Analyses of Rab8 and Rablla in Exocytic Transport from trans-Golgi Network

19: Expression and Properties of Rab7 in Endosome Function

20: Expression, Purification, and Properties of Rab8 Function in Actin Cortical Skeleton Organization and Polarized Transport

21: Properties of Rab13 Interaction with Rod cGMP Phosphodiesterase δ Subunit

22: Subcellular Localization of Rab17 by Cryo-Immunogold Electron Microscopy in Epithelial Cells Grown on Polycarbonate Filters

23: Expression and Properties of Rab25 in Polarized Madin-Darby Canine Kidney Cells

24: Purification of TRAPP from Saccharomyces cerevisiae and Identification of Its Mammalian Counterpart

Section II. ADP-Ribosylation Factor (ARF) GTPases

25: Structural and Functional Organization of ADP-Ribosylation Factor (ARF) Proteins

26: Expression and Properties of ADP-Ribosylation Factor (ARF6) in Endocytic Pathways

27: Expression and Analysis of ARNO and ARNO Mutants and Their Effects on ADP-Ribosylation Factor (ARF)-Mediated Actin Cytoskeletal Rearrangements

28: Expression, Purification, and Measurements of Activity of ARNO1, a Guanine Nucleotide Exchange Factor for ADP-Ribosylation Factor 1 (ARF1)

29: Expression, Purification, and Biochemical Properties of EFA6, a Sec7 Domain-Containing Guanine Exchange Factor for ADP-Ribosylation Factor 6 (ARF6)

30: Isolation and Properties of GRP1, an ADP-Ribosylation Factor (ARF)-Guanine Nucleotide Exchange Protein Regulated by Phosphatidylinositol 3, 4, 5-Trisphosphate

31: Functional Analysis of ADP-Ribosylation Factor (ARF) Guanine Nucleotide Exchange Factors Gea1p and Gea2p in Yeast

32: Isolation, Cloning, and Characterization of Brefeldin A-Inhibited Guanine Nucleotide-Exchange Protein for ADP-Ribosylation Factor

33: Expression, Purification, and Properties of ADP-Ribosylation Factor (ARF) GTPase Activating Protein-1

34: Expression, Analysis, and Properties of Yeast ADP-Ribosylation Factor (ARF) GTPase Activating Proteins (GAPs) Gcs1 and Glo3

35: Purification and Properties of ARD1, An ADP-Ribosylation Factor (ARF)-Related Protein with GTPase-Activating Domain

36: Purification and Characterization of GIT Family of ADP-Ribosylation Factor (ARF) GTPase-Activating Proteins

37: Assay and Purification of Phosphoinositide-Dependent ADP-Ribosylation Factor (ARF) GTPase Activating Proteins

38: Biological Properties and Measurement of Phospholipase D Activation by ADP-Ribosylation Factor (ARF)

39: Use of Aminoglycoside Antibiotics and Related Compounds to Study ADP-Ribosylation Factor (ARF) /Coatomer Function in Golgi Traffic

40: Adaptor Protein 1-Dependent Clathrin Coat Assembly on Synthetic Liposomes and Golgi Membranes

41: Receptor-Dependent Formation of COPI-Coated Vesicles from Chemically Defined Donor Liposomes

42: ADP-Ribosylation Factor (ARF) as Regulator of Spectrin Assembly at Golgi Complex

43: Purification, Properties, and Analysis of yARL3

44: Preparation and Assay of Recombinant ADP-Ribosylation Factor-Like Protein-1 (ARL1)

Section III. Sar GTPases

45: Purification and Properties of Rat Liver Sec23–Sec24 Complex

46: Purification of Functional Sec13p–Sec31p Complex, A Subunit of COPII Coat

Section IV. Dynamin GTPases

47: Expression, Purification, and Functional Assays for Self-Association of Dynamin-1

48: Analysis of Phosphoinositide Binding by Pleckstrin Homology Domain from Dynamin

49: Mapping Dynamin Interdomain Interactions with Yeast Two-Hybrid and Glutathione S-Transferase Pulldown Experiments

50: Interactions of Dynamin and Amphiphysin with Liposomes

51: Activation of Dynamin GTPase Activity by Phosphoinositides and SH3 Domain-Containing Proteins

Section V. Septin GTPases

52: Expression and Analysis of Properties of Septin CDCrel-1 in Exocytosis

Author Index

Subject Index