Dataset Information

Kinase-catalyzed Biotinylation to Map Cell Signaling Pathways (K-BMAPS): Application to Epidermal Growth Factor Signaling

ABSTRACT: Cell signaling involves a network of protein-protein interactions and post-translational modifications that govern cellular responses to environmental cues. To understand and ultimately modulate these signaling pathways to confront disease, the complex web of proteins that becomes phosphorylated after extracellular stimulation has been studied using mass spectrometry-based proteomics methods. To complement prior work and fully characterize all phosphorylated proteins after stimulation of cell signaling, we developed K-BMAPS (Kinase-catalyzed Biotinylation to MAP Signaling), which utilizes ATP-biotin as a kinase cosubstrate to biotin label substrates. As a first application of K-BMAPS, the well-characterized epidermal growth factor receptor (EGFR) kinase signaling pathway was monitored by treating EGF-stimulated HeLa lysates with ATP-biotin, following by streptavidin enrichment and quantitative mass spectrometry analysis. Based on the dynamic phosphoproteins identified, a pathway map was developed considering functional categories and known interactors of EGFR. Remarkably, 94% of K-BMAPS hit proteins were included in the EGFR pathway map. With many proteins involved in transcription, translation, cell adhesion, and GTPase signaling, K-BMAPS identified phosphoproteins associated with late and continuous signaling events. In summary, K-BMAPS is a powerful tool to map the dynamic phosphorylation governing cell signaling pathways.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: Jordan Burton

LAB HEAD: Mary Kay Pflum

PROVIDER: PXD027529 | Pride | 2021-11-03

REPOSITORIES: Pride

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Kinase-Catalyzed Biotinylation to Map Cell Signaling Pathways: Application to Epidermal Growth Factor Signaling.

Ramanayake-Mudiyanselage Vindya V Embogama D Maheeka DM Pflum Mary Kay H MKH

Journal of proteome research 20210907 10

Cell signaling involves a network of protein-protein interactions and post-translational modifications that govern cellular responses to environmental cues. To understand and ultimately modulate these signaling pathways to confront disease, the complex web of proteins that becomes phosphorylated after extracellular stimulation has been studied using mass spectrometry-based proteomics methods. To complement prior work and fully characterize all phosphorylated proteins after the stimulation of cel ...[more]

PMID: 34491762

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