Identification of Upstream Kinases by Fluorescence Complementation Mass Spectrometry.

ABSTRACT: Protein kinases and their substrates comprise extensive signaling networks that regulate many diverse cellular functions. However, methods and techniques to systematically identify kinases directly responsible for specific phosphorylation events have remained elusive. Here we describe a novel proteomic strategy termed fluorescence complementation mass spectrometry (FCMS) to identify kinase-substrate pairs in high throughput. The FCMS strategy employs a specific substrate and a kinase library, both of which are fused with fluorescence complemented protein fragments. Transient and weak kinase-substrate interactions in living cells are stabilized by the association of fluorescence protein fragments. These kinase-substrate pairs are then isolated with high specificity and are identified and qu

SUBMITTER: Zeng L 

PROVIDER: S-EPMC5658758 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature