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Global and site-specific effect of phosphorylation on protein turnover

ABSTRACT: The effects of specific modification types and sites on protein lifetime have not been illustrated in large scale. We describe a proteomic method, DeltaSILAC, to quantify the impact of site-specific phosphorylation on the turnover of thousands of proteins in live cells. Being configured on the accurate and reproducible mass spectrometry, the pulse labeling approach using stable isotope-labeled amino acids in cells (pSILAC), phosphoproteomics, as well as a novel peptide-level matching strategy, our DeltaSILAC profiling revealed a global, unexpected delaying effect of many phosphosites on protein turnover. We further found that phosphorylated sites accelerating protein turnover are functionally selected for cell fitness, enriched in Cyclin-dependent kinase substrates, and evolutionarily conserved, whereas the Glutamic acids surrounding phosphosites significantly delay protein turnover. Our investigation provides a generalizable approach and a rich resource for prioritizing the effects of phosphorylation sites on protein lifetime in the context of cell signaling and disease biology.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Rattus Norvegicus (rat) Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Yansheng Liu

LAB HEAD: Yansheng Liu

PROVIDER: PXD017496 | Pride | 2020-11-17

REPOSITORIES: Pride

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