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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.

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Includes supplementary material: sn.pub/extras

Part I Introduction 1. Concepts and Methodologies to Study Protein SUMOylation: An OverviewMichael J. Matunis and Manuel S. Rodríguez2. The Regulation of Chromatin by Dynamic SUMO ModificationsNicole R. Wilson and Mark HochstrasserPart II Procedures to Study Protein SUMOylation In Vitro3. Reconstitution of the Recombinant RanBP1 SUMO E3 Ligase ComplexTobias Ritterhoff, Hrishikesh Das, Yuqing Hao, Volkan Sakin, Annette Flotho, Andreas Werner, and Frauke Melchior4. Production and Purification of Recombinant SUMOylated Proteins Using Engineered BacteriaFrédérique Brockly, Marc Piechaczyk, and Guillaume Bossis5. A Fluorescent In Vitro Assay to Investigate Paralog Specific SUMO ConjugationNathalie Eisenhardt, Viduth K. Chaugule, and Andrea Pichler6. Identification and Characterization of SUMO-SIM InteractionsKoraljka Husnjak, Jan Keiten-Schmitz, and Stefan Müller7. Real-Time Surface Plasmon Resonance (SPR) for the Analysis of Interactions between SUMO-Traps and Mono or PolySUMO MoietiesWendy Xolalpa, Manuel S. Rodríguez, and Patrick England 8. Using Biotinylated SUMO-Traps to Analyze SUMOylated ProteinsValérie Lang, Elisa Da Silva-Ferrada, Rosa Barrio, James D. Sutherland, and Manuel S. Rodríguez9. In Vitro Characterization of Chain Depolymerization Activities of SUMO-Specific ProteasesJulia Eckhoff and R. Jürgen DohmenPart III Analysis of Protein SUMOylation Using Cell Lines10. Detection of Protein SUMOylation In Situ by Proximity Ligation AssaysUmut Sahin, Florence Jollivet, Caroline Berthier, Hugues de Thé, and Valérie Lallemand-Breitenbach<11. In Situ SUMOylation and DeSUMOylation Assays: Fluorescent Methods to Visualize SUMOylation and DeSUMOylation in Permeabilized CellsEri Yuasa and Hisato Saitoh12. Analysis of SUMOylated Proteins in Cells and In Vivo Using BioSUMO StrategyLucia Pirone, Wendy Xolalpa, Ugo Mayor, Rosa Barrio, and James D. Sutherland13. Label-Free Identification and Quantification of SUMO Target ProteinsIvo A. Hendriks and Alfred C.O. Vertegaal 14. The Use of Multimeric Protein Scaffolds for Identifying Multi-SUMO Binding ProteinsElisa Aguilar-Martínez and Andrew D. Sharrocks15. Isolation of In Vivo SUMOylated Chromatin-Bound ProteinsTasneem Bawa-Khalfe


Titel: SUMO
Untertitel: Methods and Protocols
EAN: 9781493963560
ISBN: 978-1-4939-6356-0
Format: Fester Einband
Herausgeber: Springer, Berlin
Genre: Biologie
Anzahl Seiten: 306
Größe: H25mm x B263mm x T182mm
Jahr: 2016
Untertitel: Englisch
Auflage: 1st ed. 2016

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