SUMO Protocols -

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Sommaire:
Part I. Introduction 1 The SUMO System: An Overview Helle D. Ulrich Part II. Identification of SUMO Targets 2 Identification of SUMO Target Proteins by Quantitative Proteomics Jens S. Andersen, Ivan Matic and Alfred C. O. Vertegaal 3 Identification of SUMO-Conjugated Proteins and their SUMO Attachment Sites Using Proteomic Mass Spectrometry James A. Wohlschlegel 4 Identification of SUMO Targets through In Vitro Expression Cloning Christian B. Gocke and Hongtao Yu 5 Enhanced Detection of In Vivo SUMO Conjugation by Ubc9 Fusion-Dependent Sumoylation (UFDS) Rainer Niedenthal 6 In Vivo Detection and Characterization of Sumoylation Targets in Saccharomyces cerevisiae Helle D. Ulrich and Adelina A. Davies Part III. Identification of SUMO Interactors 7 Identification of SUMO-Interacting Proteins by Yeast Two-Hybrid Analysis Mary B. Kroetz and Mark Hochstrasser 8 Identification of SUMO Binding Motifs by NMR Candace S. Seu and Yuan Chen Part IV. Functional Analysis of Sumoylation In Vivo 9 Regulation of Transcription Factor Activity by SUMO Modification Jian Ouyang, Alvaro Valin and Grace Gill 10 Characterization of the Effects and Functions of Sumoylation Through Rapamycin-Mediated Heterodimerization Shanshan Zhu and Michael J. Matunis Part V. Biochemical Studies of SUMO Conjugation 11 Purification of SUMO Conjugating Enzymes and Kinetic Analysis of Substrate Conjugation Ali A. Yunus and Christopher D. Lima 12 Performing In Vitro Sumoylation Reactions Using Recombinant Enzymes Andreas Werner, Marie-Christine Moutty, Ulrike M?ller and Frauke Melchior 13 Preparation of Sumoylated Substrates for Biochemical Analysis Puck Knipscheer, Helene Klug, Titia K.Sixma and Andrea Pichler 14 Strategies for the Expression of SUMO-Modified Target Proteins in Escherichia coli Hisato Saitoh, Junsuke Uwada and Azusa Kawasaki Part VI. Biochemical Studies of SUMO Deconjugation 15 Preparation of SUMO Proteases and Kinetic Analysis Using Endogenous Substrates David Reverter and Christopher D. Lima 16 An In Vitro FRET-Based Assay for the Analysis of SUMO Conjugation and Isopeptidase Cleavage Nicolas Stankovic-Valentin, Lukasz Kozaczkiewicz, Katja Curth and Frauke Melchior 17 FRET-Based In Vitro Assays for the Analysis of SUMO Protease Activities Michael H. Tatham and Ronald T. Hay 18 Detection and Characterization of SUMO Protease Activity Using a Sensitive Enzyme-Based Reporter Assay Craig A. Leach, Xufan Tian, Michael R. Mattern and Benjamin Nicholson Part VII. Manipulation and Alternative Uses of the SUMO System 19 Inhibition of the SUMO Pathway by Gam1 Mariaelena Pozzebon, Chiara V. Segre and Susanna Chiocca 20 SUMO Fusion Technology for Enhanced Protein Production in Prokaryotic and Eukaryotic Expression Systems Tadas Panavas, Carsten Sanders and Tauseef R. Butt