Exocytosis and Endocytosis

ISBN-13: 9781493946945 / Angielski / Miękka / 2016 / 435 str.

Andrei I. Ivanov

Opis

Focused on one of the most important and fascinating fields of modern biology, Exocytosis and Endocytosis, Second Edition presents a comprehensive collection of protocols for cutting edge, recently developed methods of studying exocytosis and endocytosis in vitro and in vivo. Divided into four easy-to-follow sections, chapters focus on diverse research topics including cell-free and biochemical assays of exocytosis and endocytosis, probing protein, membrane and organelle trafficking in cultured cells and powerful in vivo imaging approaches to investigate different physiological processes involving exocytosis and endocytosis. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.Highly practical and clearly written, Exocytosis and Endocytosis, Second Edition will serve both seasoned researchers as well as newcomers to the field and will provide a unique resource and expert guidance to modern laboratory techniques developed for examining protein and membrane trafficking in simple cellular systems and model organisms.

Spis treści

Part I. An Overview of Methods for Analyzing Vesicular Trafficking

 

1. Pharmacological Inhibitors of Exocytosis and Endocytosis:  Novel Bullets for Old Targets

Andrei I. Ivanov

 

2. Systematic Analysis of Endocytosis by Cellular Perturbations

Lena Kühling and Mario Schelhaas

 

Part II. Cell-Free and Biochemical Assays of Exocytosis and Endocytosis

 

3. Real Time Detection of SNARE Complexes with FRET Using the Tetracysteine System

Oleg Varlamov

 

4. Profiling Lysine Ubiquitination by Selective Enrichment of Ubiquitin Remnant-Containing Peptides

Guoqiang Xu, Alessia Deglincerti, Jeremy S. Paige, and Samie R. Jaffrey

 

5. Secretion of Circular Proteins Using Sortase

Karin Strijbis and Hidde Ploegh

 

6. Fractionation of Subcellular Membrane Vesicles of Epithelial and Non-Epithelial Cells by OptiPrepTM Density Gradient Ultracentrifugation

Xuhang Li and Mark Donowitz

 

7. Combining Pulsed SILAC Labeling and Click-Chemistry for Quantitative Secretome Analysis

Katrin Eichelbaum and Jeroen Krijgsveld

 

Part III. Imaging and Ultrastructural Analysis of Vesicle Trafficking in Cultured Cells

 

8. Probabilistic Density Maps to Study the Spatial Organization of Endocytosis

Jean-Philippe Grossier, Bruno Goud, and Kristine Schauer

 

9. Use of Kaede and Kikume Green-Red Fusions for Live Cell Imaging of G Protein-Coupled Receptors

Antje Schmidt, Burkhard Wiesner, Ralf Schulein, and Anke Teichmann

 

10. Halo Tag as a Tool to Investigate Peroxisome Dynamics in Cultured Mammalian Cells

Marc Fransen

 

11. SNAP-Tag to Monitor Trafficking of  Membrane Proteins in Polarized Epithelial Cells

Emily H. Stoops, Glen A. Farr, Michael Hull, and Michael J. Caplan

 

12. FLAsH-PALM:  Super-Resolution Pointillist Imaging with FLAsH-tetracysteine Labeling

Mickael Lelek, Francesca Di Nunzio, and Christophe Zimmer

 

13. Analysis of Protein Dynamics with Tandem Fluorescent Protein Timers

Anton Khmelinskii and Michael Knop

 

14. Synchronization of Secretory Cargo Trafficking in Populations of Cells

Gaelle Boncompain and Franck Perez

 

15. Use of Transmembrane FRET to Investigate the Internalization of Glycosylated Proteins

Yoshimi Haga and Tadashi Suzuki

 

16. A Method to Rapidly Induce Organelle-specific Molecular Activities and Membrane Tethering

Toru Komatsu and Takanari Inoue

 

17. A Novel Pair of Split Venus Fragments to Detect Protein-Protein Interactions by In Vitro and In Vivo Biomolecular Fluorescence Complementation Assays

Kazumasa Ohashi and Kensaku Mizuno

 

18. Real Time Investigation of Plasma Membrane Deformation and Fusion Pore Expansion Using Polarized Total Internal Reflection Fluorescence Microscopy

Daniel R. Passmore, Tejeshwar Rao, and Arun Anantharam

 

19. Nanocones to Study Initial Steps of Endocytosis

Sangmoo Jeong and Milos Galic

 

20. A Novel Permeabilization Protocol to Obtain Intracellular 3D Immunolabeling for Electron Tomography

Nuria Jimenez and Jan A. Post 

 

21. VIS2FIX: Rapid Chemical Fixation of Vitreous Sections for Immuno-Electron Microscopy

Matthia A. Karreman and Elly G. Van Donselaar

 

Part IV.  Analysis of Exocytosis and Endocytosis in Model Organisms

 

22. Chemical Genomics: Characterizing Target Pathways for Bioactive Compounds Using the Endomembrane Trafficking Network

Cecilia Rodriguez-Furlán, Glenn R. Hicks, and Lorena Norambuena

 

23. Application of RNAi Technology and Fluorescent Protein Markers to Study Membrane Traffic in C. elegans

Jachen A. Solinger, Dmitry Poteryaev, and Anne Spang

 

24. Visualization of Clathrin-Mediated Endocytosis in Live Drosophila Egg Chambers

Anupma Jha and Linton M. Traub

 

25. Novel Extraction Protocol to Probe the Role of Cholesterol in Synaptic Vesicle Recycling

Jeffrey S. Dason and Milton P. Charlton

 

26. Microfluidic Devices for Imaging Trafficking Events In Vivo Using Genetic Model Organisms

Sudip Mondal and Sandhya P. Koushika

 

27. The “In Situ” Proximity Ligation Assay to Probe Protein-Protein Interactions in Intact Tissues

Arianna Bellucci, Chiara Fiorentini, Michela Zaltieri, Cristina Missale, and PierFranco Spano

 

28. Probing the Role of the Actin Cytoskeleton During Regulated Exocytosis by Intravital Microscopy

Oleg Milberg, Muhibullah Tora, Akiko Shitara, Taishin Takuma, Andrius Masedunskas, and Roberto Weigert

 

29. Measurement of Dynamic F-actin Changes During Exocytosis

Peter Thorn