While the small ubiquitin-related modifier SUMO uses a conjugation and de-conjugation system closely related to that of ubiquitin itself, its functions arehighly 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 SUMOresearch. The first half of the volume focuses on the identification and functional characterization of SUMO targets and interactors, while the second halfcovers 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. First methods collection devoted entirely to the SUMO system Provides comprehensive practical advice from leading experts in the field Covers a range of complementary approaches for SUMO target identification Presents step-by-step protocols for the purification andmanipulation of SUMO conjugation and de-conjugation enzymes Includes a description of SUMO fusion technology for improved recombinant protein production INDICE: From the contents The SUMO System: An Overview.- Identification ofSUMO 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-Interacting Proteins by YeastTwo-Hybrid Analysis.- Identification of SUMO Binding Motifs by NMR.- Regulation of Transcription Factor Activity by SUMO Modification.- Characterization ofthe Effects and Functions of Sumoylation Through Rapamycin-Mediated Heterodimerization.- 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.
- ISBN: 978-1-934115-80-0
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 310
- Fecha Publicación: 01/11/2008
- Nº Volúmenes: 1
- Idioma: Inglés