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Large-Scale Identification of the Arginine Methylome by Mass Spectrometry

Publikation: Bidrag til bog/antologi/rapportBidrag til bog/antologiForskningfagfællebedømt

The attachment of one or more methylation groups to the side chain of arginine residues is a regulatory mechanism for cellular proteins. Recent advances in mass spectrometry-based characterization allow comprehensive identification of arginine methylation sites by peptide-level enrichment strategies. Described in this unit is a 4-day protocol for enrichment of arginine-methylated peptides and subsequent identification of thousands of distinct sites by mass spectrometry. Specifically, the protocol explains step-by-step sample preparation, enrichment using commercially available antibodies, prefractionation using strong cation exchange, and identification using liquid chromatography coupled to tandem mass spectrometry. A strategy for relative quantification is described using stable isotope labeling by amino acids in cell culture (SILAC). Approaches for analysis of arginine methylation site occupancy are also discussed. Collectively, the unit describes the essential parameters required for a successful and comprehensive experiment detailing the arginine methylome. © 2015 by John Wiley & Sons, Inc.

OriginalsprogEngelsk
TitelCurrent Protocols in Protein Science
Antal sider17
Vol/bind82
Publikationsdato2 nov. 2015
Kapitel24.7
ISBN (Elektronisk) 9780471140863
DOI
StatusUdgivet - 2 nov. 2015
NavnCurrent Protocols in Protein Science
ISSN1934-3655

ID: 176889772