SUMO protocols / edited by Helle D. Ulrich.

Includes bibliographical references and index.

The SUMO system : an overview / Helle D. Ulrich -- Identification of SUMO target proteins by quantitative proteomics / Jens S. Anderson, Ivan Matic, and Alfred C.O. Vertegaal -- Identification of SUMO-conjugated proteins and their SUMO attachment sites using proteomic mass spectrometry / James A. Wohlschlegel -- Identification of SUMO targets through in vitro expression cloning / Christian B. Gocke and Hongtao Yu -- Enhanced detection of in vivo SUMO conjugation by Ubc9 fusion-dependent sumoylation (UFDS) / Rainer Niedenthal -- In vivo detection and characterization of sumoylation targets in Saccharomyces cervisiae / Helle D. Ulrich and Adelina A. Davies -- Identification of SUMO-interacting proteins by yeast two-hybrid analysis / Mary B. Kroetz and Mark Hochstrasser -- Identification of SUMO-binding motifs by NMR / Candace S. Seu and Yuan Chen -- Regulation of transcription factor activity by SUMO modification / Jian Ouyang, Alvaro Valin, and Grace Gill -- Characterization of the effects and functions of sumoylation through rapamycin-mediated heterodimerization / Shanshan Zu and Michael J. Matunis -- Purification of SUMO conjugating enzymes and kinetic analysis of substrate conjugation / Ali A. Yunus and Christopher D. Lima -- Performing in vitro sumoylation reactions using recombinant enzymes / Andreas Werner [and others] -- Preparation of sumoylated substrates for biochemical analysis / Puck Knipscheer [and others] -- Strategies for the expression of SUMO-modified target proteins in Escherichia coli / Hisato Saitoh, Junsuke Uwada, and Azusa Kawasaki -- Preparation of SUMO proteases and kinetic analysis using endogenous substrates / David Reverter and Christopher D. Lima -- An in vitro FRET-based assay for the analysis of SUMO conjugation and isopeptidase cleavage / Nicolas Stankovic-Valentin [and others] -- FRET-based in vitro assays for the analysis of SUMO protease activities / Michael H. Tatham and Ronald T. Hay --Detection and characterization of SUMO protease activity using a sensitive enzyme-based reporter assay / Craig A. Leach [and others] -- Inhibition of the SUMO pathway by Gam1 / Mariaelena Pozzebon, Chiara V. Segré, and Susanna Chiocca -- SUMO fusion technology for enhanced protein production in prokaryotic and eukaryotic expression systems.

Print version record.

While the small ubiquitin-related modifier SUMO uses a conjugation and de-conjugation system closely related to that of ubiquitin itself, its functions are highly diverse and largely independent of the ubiquitin system. In SUMO Protocols, leading experts present a collection of methods intended to highlight important features unique to the biology of SUMO in order to support future SUMO research. The first half of the volume focuses on the identification and functional characterization of SUMO targets and interactors, while the second half covers essential biochemical methods for the manipulation of the enzymatic components of the SUMO system. Written in the highly successful Methods in Molecular Biology series format, the chapters include brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and the Notes sections, which provide tips on troubleshooting and avoiding known pitfalls. Unique and cutting-edge, SUMO Protocols provides the necessary tools for biochemists, molecular and cellular biologists as well as research-oriented clinicians not yet familiar with this vital system.