Main description:

This second edition volume expands on the previous edition with a discussion of new research and discoveries in the Rab field. Chapters in this book cover topics such as new information on Rab regulation and localization; interaction; function; and diseases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Rab GTPases: Methods and Protocols, Second Edition is a valuable resource for scientists working in the fields of Rab and other small GTPases, and beyond.

Contents:

Preface...
Table of Contents...
List of Reviewers...
Contributing Authors...

1. Newer Methods Drive Recent Insights into Rab GTPase Biology: An Overview
Guangpu Li and Nava Segev

2. Rab29 Fast Exchange Mutants: Characterization of a Challenging Rab GTPase
Rachel C. Gomez, Edmundo G. Vides, and Suzanne R. Pfeffer

3. High-Throughput Assay for Profiling the Substrate Specificity of Rab GTPase-Activating Proteins
Ashwini K. Mishra and David G. Lambright

4. Detecting Endogenous Rab8 Activation
Samuel J. Tong, Richard M. Lucas, Zhijian Xiao, Lin Luo, and Jennifer L. Stow

5. Testing the Phenotypic Effects of a Rab Chimera that Resolves Exchange Factor Specificity from Effector Specificity
Hua Yuan and Peter Novick

6. Profiling Structural Alterations during Rab5 Nucleotide Exchange by HDX-MS
Janelle Lauer and Marino Zerial

7. CLEM Characterization of Rab8 and Associated Membrane Trafficking Regulators at Primary Cilium Structures
Quanlong Lu and Christopher J. Westlake

8. Imaging of Spatial Cycling of Rab GTPase in the Cell
Fu Li and Yao-Wen Wu

9. Deconvolution of Multiple Rab Binding Domains using the Batch Yeast 2-Hybrid Method DEEPN
Tabitha A. Peterson and Robert C. Piper

10. Determination of the Rab27-Effector Binding Affinity Using a High-Throughput FRET-Based Assay
Raghdan Z. Al-Saad, Ian Kerr, and Alistair N. Hume

11. Methods to Study the Unique SOCS Box Domain of the Rab40 Small GTPase Subfamily
Emily D. Duncan, Ezra Lencer, Erik Linklater, and Rytis Prekeris

12. Using GBP Nanotrap to Restore Autophagy in the Rab5/Vps21 Mutant by Forcing Snf7 and Atg17 Interaction
Mengzhu Zhao and Yongheng Liang

13. Establishing Regulation of a Dynamic Process by Ypt/Rab GTPase: A Case for Cisternal Progression
Jane J. Kim, Zanna Lipatova, and Nava Segev

14. Methods for Assessing the Regulation of a Kinase by the Rab GTPase Ypt1
Juan Wang, Shensen Wang, and Susan Ferro-Novick

15. Qualitative and Quantitative Assessment of the Role of Endocytic Regulatory and/or Rab Proteins in Mitochondrial Fusion and Fission
Trey Farmer and Steve Caplan

16. Characterization of the Role of Rab18 in Mediating LD-ER Contact and LD Growth
Dijin Xu, Peng Li, and Li Xu

17. Methods for Establishing Rab Knockout MDCK Cells
Riko Kinoshita, Yuta Homma, and Mitsunori Fukuda

18. Generating Rab6 Conditional Knockout Mice
Sabine Bardin and Bruno Goud

19. Use of Immunohistochemistry to Determine Expression of Rab5 Subfamily of GTPases in Mature and Developmental Brains
Kwok-Ling Kam, Paige Parrack, Marcellus Banworth, Sheeja Aravindan, Guangpu Li, and Kar-Ming Fung

20. Assessing Rab5 Activation in Health and Disease
Anna Pensalfini, Ying Jiang, Seonil Kim, and Ralph A. Nixon

21. Quantitative Fluorescence Microscopy for Detecting Mammalian Rab Vesicles within the Parasitophorous Vacuole of the Human Pathogen Toxoplasma gondii
Julia D. Romano, Eric J. Hartman, and Isabelle Coppens

Subject Index List...