Motif-centric phosphoproteomics to target kinase-mediated signaling pathways.

ABSTRACT: Identifying cellular phosphorylation pathways based on kinase-substrate relationships is a critical step to understanding the regulation of physiological functions in cells. Mass spectrometry-based phosphoproteomics workflows have made it possible to comprehensively collect information on individual phosphorylation sites in a variety of samples. However, there is still no generic approach to uncover phosphorylation networks based on kinase-substrate relationships in rare cell populations. Here, we describe a motif-centric phosphoproteomics approach combined with multiplexed isobaric labeling, in which in vitro kinase reactions are used to generate targeted phosphopeptides, which are spiked into one of the isobaric channels to increase detectability. Proof-of-concept experiments demo

SUBMITTER: Tsai CF 

PROVIDER: S-EPMC9017188 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature