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Covalent inhibition of the peptidyl-prolyl isomerase Pin1 by Sulfopin results in a broad impact on the phosphoproteome of human osteosarcoma U2-OS cells

ABSTRACT: Peptidyl-prolyl isomerase, NIMA-interacting protein 1 (Pin1) catalyzes the cis-trans interconversion of the inflexible bond between serine or threonine residues and proline at the +1 position (pSer/pThr-Pro). While initially discovered as an essential regulator of cell division, Pin1 has since been identified as a regulator of many biological processes and is associated with numerous malignancies and neurodegenerative disorders. Pin1 has been shown to influence phosphorylation by modulating phosphatase accessibility. However, it can also indirectly regulate phosphorylation by isomerizing peptidyl-prolyl bonds on kinases, affecting their subcellular localization and/or substrate specificity. Here, SILAC-based mass spectrometry was employed to identify proteomic and phosphoproteomic changes in human osteosarcoma (U2-OS) cells in response to treatment with the selective covalent Pin1 inhibitor Sulfopin. We confirmed that Sulfopin covalently binds Pin1 and profiled Pin1-dependent changes to the proteome and phosphoproteome, identifying 803 phosphosites that underwent significant Sulfopin-dependent changes. The identified phosphosites include substrates for a number of distinct kinases, including AKT1, AURKA, CDK1, and CK2. Overall, this study reveals broad impact of Sulfopin on the phosphoproteome improving our understanding of how Pin1 modulates complex regulatory kinase networks in living cells.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Tibia, U2-os Cell, Epithelial Cell, Cell Culture

DISEASE(S): Osteosarcoma

SUBMITTER: Scott Roffey

LAB HEAD: David Litchfield

PROVIDER: PXD049431 | Pride | 2025-06-24

REPOSITORIES: Pride

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