Selective Enrichment and Quantitation of Phosphoproteins and Phosphopeptides Involved in Cell Proliferation
John Rogers1
1 Thermo Fisher Scientific
The serine/threonine kinase Akt regulates multiple key proteins involved in apoptosis (Bad), cell proliferation (FOXO), translation (mTOR and its effector proteins), and glycogen synthesis (GSK3b). To better understand the complex regulation of phosphorylation in cellular proliferation, phosphoproteins and phosphopeptides were enriched from insulin-responsive 293T cells treated with or without wortmannin and rapamycin. Phosphoproteomes were quantitatively analysed with TMT isobaric mass tags , interpreted by pathway analysis, and verified by Western blot analysis. This analysis enabled the characterization of novel effector proteins and phosphorylation events involved in cellular proliferation.