SUMO Protocols
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 Expre...
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| Main Author: | |
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| Format: | Edited Volume |
| Language: | English |
| Published: |
Totowa, NJ
Humana Press
2009
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| Series: | Methods in molecular biology
497 |
| In: |
Methods in molecular biology (497)
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| Volumes / Articles: | Show Volumes / Articles. |
| DOI: | 10.1007/978-1-59745-566-4 |
| Subjects: | |
| Online Access: | Resolving-System, lizenzpflichtig: https://doi.org/10.1007/978-1-59745-566-4 Cover: https://swbplus.bsz-bw.de/bsz285882457cov.jpg 
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| Author Notes: | edited by Helle D. Ulrich |
| Summary: | 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. 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. |
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| Physical Description: | Online Resource |
| ISBN: | 9781597455664 |
| DOI: | 10.1007/978-1-59745-566-4 |