Incremental Modification in the Existing Approaches for Affinity Chromatographic Enrichment of Phosphoproteins Improves Their Profile in Liquid Chromatography-Tandem Mass Spectrometry Analysis.

ABSTRACT: Cell signalling is a vital process in cell physiology, which is driven by protein phosphorylation. Global phosphoproteome analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) has thus gained importance in cell signalling research. However, phosphoprotein identification by LC-MS/MS in whole cell lysates, which are complex protein mixtures, is hindered by their poor ionization coupled with suppression of peaks due to low abundance. Enrichment by immobilized metal ion- and metal oxide-affinity chromatography (IMAC and MOAC), which preferentially enrich multi- and mono-phosphorylated proteins, respectively, have improved their detection by MS. However, preferential enrichment limits phosphoproteome coverage in global analyses of cell lysates which contain mono- and multi-phosp

SUBMITTER: Agrawal N 

PROVIDER: S-EPMC11789765 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature