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Direct mapping of ligandable tyrosines and lysines in cells with chiral sulfonyl fluoride probes

ABSTRACT: Advances in chemoproteomic technology have revealed covalent interactions between small molecules and protein nucleophiles, primarily cysteine, on a proteome-wide scale. Most chemoproteomic screening approaches are indirect, relying on competition between electrophilic fragments and a minimalist electrophilic probe with inherently limited proteome coverage. Here, we develop a chemoproteomic platform for direct electrophile-site identification based on enantiomeric pairs of clickable arylsulfonyl fluoride probes. Using stereoselective site modification as a proxy for ligandability in intact cells, we identified 634 tyrosines and lysines within functionally diverse protein sites, liganded by structurally diverse probes. Among multiple validated sites, we discovered a chiral probe that modifies Y228 in the MYC binding site of the epigenetic regulator WDR5, as revealed by a high-resolution crystal structure. A distinct chiral probe stimulates tumor cell phagocytosis by covalently modifying Y387 in the recently discovered immuno-oncology target, APMAP. Our work provides a deep resource of ligandable tyrosines and lysines for the development of covalent chemical probes.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): T Cell, Cell Culture

DISEASE(S): Acute Leukemia

SUBMITTER: Ying Chen

LAB HEAD: Jack Taunton

PROVIDER: PXD042307 | Pride | 2023-05-18

REPOSITORIES: Pride

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