Sumo: Methods and Protocols

ISBN-13: 9781493963560 / Angielski / Twarda / 2016 / 306 str.

Manuel S. Rodriguez

Spis treści

Part I Introduction

1. Concepts and Methodologies to Study Protein SUMOylation: An Overview
Michael J. Matunis and Manuel S. Rodríguez

2. The Regulation of Chromatin by Dynamic SUMO Modifications
Nicole R. Wilson and Mark Hochstrasser

Part II Procedures to Study Protein SUMOylation In Vitro

3. Reconstitution of the Recombinant RanBP1 SUMO E3 Ligase Complex
Tobias Ritterhoff, Hrishikesh Das, Yuqing Hao, Volkan Sakin, Annette Flotho, Andreas Werner, and Frauke Melchior

4. Production and Purification of Recombinant SUMOylated Proteins Using Engineered Bacteria

Frédérique Brockly, Marc Piechaczyk, and Guillaume Bossis

5. A Fluorescent In Vitro Assay to Investigate Paralog Specific SUMO Conjugation

Nathalie Eisenhardt, Viduth K. Chaugule, and Andrea Pichler

6. Identification and Characterization of SUMO-SIM Interactions

Koraljka Husnjak, Jan Keiten-Schmitz, and Stefan Müller

7. Real-Time Surface Plasmon Resonance (SPR) for the Analysis of Interactions between SUMO-Traps and Mono or PolySUMO Moieties

Wendy Xolalpa, Manuel S. Rodríguez, and Patrick England 

8. Using Biotinylated SUMO-Traps to Analyze SUMOylated Proteins

Valérie Lang, Elisa Da Silva-Ferrada, Rosa Barrio, James D. Sutherland, and Manuel S. Rodríguez

9. In Vitro Characterization of Chain Depolymerization Activities of SUMO-Specific Proteases

Julia Eckhoff and R. Jürgen Dohmen

Part III Analysis of Protein SUMOylation Using Cell Lines

10. Detection of Protein SUMOylation In Situ by Proximity Ligation Assays

Umut Sahin, Florence Jollivet, Caroline Berthier, Hugues de Thé, and Valérie Lallemand-Breitenbach

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11. In Situ SUMOylation and DeSUMOylation Assays: Fluorescent Methods to Visualize SUMOylation and DeSUMOylation in Permeabilized Cells

Eri Yuasa and Hisato Saitoh

12. Analysis of SUMOylated Proteins in Cells and In Vivo Using BioSUMO Strategy

Lucia Pirone, Wendy Xolalpa, Ugo Mayor, Rosa Barrio, and James D. Sutherland

13. Label-Free Identification and Quantification of SUMO Target Proteins

Ivo A. Hendriks and Alfred C.O. Vertegaal 

14. The Use of Multimeric Protein Scaffolds for Identifying Multi-SUMO Binding Proteins

Elisa Aguilar-Martínez and Andrew D. Sharrocks

15. Isolation of In Vivo SUMOylated Chromatin-Bound Proteins

Tasneem Bawa-Khalfe

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Part IV Procedures to Study Protein SUMOylation In Vivo<

16. Identification of Substrates of Protein-Group SUMOylation

Ivan Psakhye and Stefan Jentsch

17. Tools to Study SUMO Conjugation in Caenorhabditis elegans

Federico Pelisch and Ronald T. Hay

18. Purification of SUMO Conjugates from Arabidopsis for Mass Spectrometry Analysis

Thérèse C. Rytz, Marcus J. Miller, and Richard D. Vierstra

19. Detection of SUMOylation in Plasmodium falciparum

Katherine H. Reiter and Michael J. Matunis

20. Systematic Localization and Identification of SUMOylation Substrates in Knock-In Mice Expressing Affinity-Tagged SUMO1

Marilyn Tirard and Nils Brose 

This volume explores various methodologies to study biochemical, molecular, and cellular biology aspects of some processes regulated by protein SUMOylation. SUMO: Methods and Protocols is organized into four parts, and starts with an historical overview on protein SUMOylation and a presentation of the methods included in the book. The first part also includes a review on chromatin regulation by dynamic SUMO modifications. The second part focuses on in vitro techniques, including biochemical methods to study mechanistic aspects of protein SUMOylation. The third part includes protocols to be used with cell cultures, which often are the first approaches used in most laboratories. The final part includes methodologies adapted for the analysis in vivo using distinct model organisms. Written in the highly successful Methods in Molecular Biology series format, chapters include a brief introduction to the subject, a list of necessary materials and reagents, a step-by-step reproducible laboratory protocol ending with a Notes section on troubleshooting tips, and tips and strategies to avoid known pitfalls.

Unique and cutting-edge, SUMO: Methods and Protocols provides a comprehensive source of protocols for specialists and researchers not familiar with this vital system.

Blurb

This volume explores various methodologies to study biochemical, molecular, and cellular biology aspects of some processes regulated by protein SUMOylation. SUMO: Methods and Protocols is organized into four parts, and starts with an historical overview on protein SUMOylation and a presentation of the methods included in the book. The first part also includes a review on chromatin regulation by dynamic SUMO modifications. The second part focuses on in vitro techniques, including biochemical methods to study mechanistic aspects of protein SUMOylation. The third part includes protocols to be used with cell cultures, which often are the first approaches used in most laboratories. The final part includes methodologies adapted for the analysis in vivo using distinct model organisms. Written in the highly successful Methods in Molecular Biology series format, chapters include a brief introduction to the subject, a list of necessary materials and reagents, a step-by-step reproducible laboratory protocol ending with a Notes section on troubleshooting tips, and tips and strategies to avoid known pitfalls.

Unique and cutting-edge, SUMO: Methods and Protocols provides a comprehensive source of protocols for specialists and researchers not familiar with this vital system.