SUMO Protocols / Edition 1

by Helle Ulrich

Overview

Post-translational protein modifications by members of the ubiquitin family are widely recognized as important regulatory control systems for a variety of biological pathways. Their influence on eukaryotic cellular metabolism is comparable to that of other modifi- tions such as phosphorylation, acetylation and methylation. The small ubiquitin-related modifier SUMO uses a conjugation and de-conjugation system closely related to that of ubiquitin itself; yet, the functions of the SUMO system are highly diverse and largely independent of the ubiquitin system. SUMO modification controls the activity of tr- scription factors and can influence protein stability, but it also contributes to nucleo-cy- plasmic transport, chromosome segregation and DNA damage repair. As a consequence, the SUMO system pervades virtually all areas of basic molecular and cell biology, and scientists from different backgrounds, including medical researchers, are likely to enco- ter SUMO in the course of their studies. This volume, SUMO Prools, therefore aims at presenting a collection of methods relevant to SUMO research in order to make these tools available to biochemists, molecular and cell biologists as well as research-oriented clinicians not yet familiar with the system. In contrast to the ubiquitin system, which has been the subject of several reviews and methods collections, no practical compendium entirely devoted to SUMO has been p- lished.

Product Details

ISBN-13:	9781617379451
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/19/2010
Series:	Methods in Molecular Biology , #497
Edition description:	Softcover reprint of hardcover 1st ed. 2009
Pages:	324
Product dimensions:	7.60(w) x 10.24(h) x 0.04(d)

Table of Contents

The SUMO System: An Overview.- Identification of Sumo Targets.- Identification of SUMO Target Proteins by Quantitative Proteomics.- Identification of SUMO-Conjugated Proteins and their SUMO Attachment Sites Using Proteomic Mass Spectrometry.- Identification of SUMO Targets Through In Vitro Expression Cloning.- Enhanced Detection of In Vivo SUMO Conjugation by Ubc9 Fusion-Dependent Sumoylation (UFDS).- In Vivo Detection and Characterization of Sumoylation Targets in Saccharomyces cerevisiae.- Identification Of Sumo Interactors.- Identification of SUMO-Interacting Proteins by Yeast Two-Hybrid Analysis.- Identification of SUMO-Binding Motifs by NMR.- Functional Analysis Of Sumoylation In Vivo.- Regulation of Transcription Factor Activity by SUMO Modification.- Characterization of the Effects and Functions of Sumoylation Through Rapamycin-Mediated Heterodimerization.- Biochemical Studies Of Sumo Conjugation.- Purification of SUMO Conjugating Enzymes and Kinetic Analysis of Substrate Conjugation.- Performing In Vitro Sumoylation Reactions Using Recombinant Enzymes.- Preparation of Sumoylated Substrates for Biochemical Analysis.- Strategies for the Expression of SUMO-Modified Target Proteins in Escherichia coli.- Biochemical Studies of Sumo Deconjugation.- Preparation of SUMO Proteases and Kinetic Analysis Using Endogenous Substrates.- An In Vitro FRET-Based Assay for the Analysis of SUMO Conjugation and Isopeptidase Cleavage.- FRET-Based In Vitro Assays for the Analysis of SUMO Protease Activities.- Detection and Characterization of SUMO Protease Activity Using a Sensitive Enzyme-Based Reporter Assay.- Manipulation And Alternative Uses Of The Sumo System.- Inhibition of the SUMO Pathway by Gam1.- SUMO Fusion Technology for Enhanced Protein Production in Prokaryotic and Eukaryotic Expression Systems.

Editorial Reviews

From the reviews:

"The modification of proteins by small ubiquitin-like modification (SUMO) is relevant to virtually all areas of molecular and cell biology. SUMO Prools is a quite comprehensive volume describing various in vivo and in vitro techniques … . The target audience is likely to be postdoctoral fellows and PhD students in laboratories not only dedicated to SUMO pathways but those beginning their initial forays in the field … and regulated by SUMO. … for those pursuing SUMO-specific modification this is a worthwhile addition to their bookshelves." (Peter O’Hare, Microbiology Today, July, 2009)

From the Publisher