SILAC for global phosphoproteomic analysis.
Establishing the phosphorylation pattern of proteins in a comprehensive fashion is an important goal of a majority of cell signaling projects. Phosphoproteomic strategies should be designed in such a manner as to identify sites of phosphorylation as well as to provide quantitative information about the extent of phosphorylation at the sites. In this chapter, we describe an experimental strategy that outlines such an approach using stable isotope labeling with amino acids in cell culture (SILAC) coupled to LC-MS/MS. We highlight the importance of quantitative strategies in signal transduction as a platform for a systematic and global elucidation of biological processes.
Pimienta
Genaro
G
Department of Biological Chemistry, McKusick-Nathans Institute of Genetic Medicine, Baltimore, MD, USA.
Chaerkady
Raghothama
R
Pandey
Akhilesh
A
D000465:Algorithms; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D010750:Phosphoproteins / Q000032:analysis* / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000378:metabolism
D010750:Phosphoproteins; D020543:Proteome