Sequential gel-assisted digestion for concomitant analysis of phosphorylated and glycosylated membrane proteome

摘要

Alterations in glycan and phosphorylation profiles of membrane proteins have been correlated with certain disease state. More importantly, 90percent of therapeutics protein drugs are glycosylated membrane proteins; phosphorylation is generally considered the most direct evidences for the perturbed signaling pathway in disease. However, it is still a great challenge to analyze the glycosylated or phosphorylated membrane proteins due to the facts that membrane proteins are highly hydrophobic, low abundant and bear heterogeneous glycosylation and phosphorylation. To facilitate sensitive and efficient characterization to analyze glycosylated and phosphorylated membrane proteins, we proposed that de-glycosylation in combination with our newly developed gel-assisted digestion not only enhanced membrane protein detection but also provide concomitant analysis of membrane proteins and their glycosylation and phosphorylation sites.