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Ogasawara et al_photo-stereoprobes

ABSTRACT: Chemical proteomics enables the global assessment of small molecule-protein interactions in native biological systems and has emerged as a versatile approach for ligand discovery. The range of small molecules explored by chemical proteomics has, however, been limited. Here, we describe a diversity-oriented synthesis (DOS)-inspired library of stereochemically-defined compounds bearing diazirine and alkyne units for UV light-induced covalent modification and click chemistry enrichment of interacting proteins, respectively. We find that these ‘photo-stereoprobes’ interact in a stereoselective manner with hundreds of proteins from various structural and functional classes in human cells and demonstrate that these interactions can form the basis for high-throughput screening-compatible nanoBRET assays. Integrated phenotypic analysis and chemical proteomics identified photo-stereoprobes that modulate autophagy by engaging the mitochondrial serine protease CLPP. Our findings show the utility of photo-stereoprobes for expanding the ligandable proteome, furnishing target engagement assays, and discovering and characterizing bioactive small molecules by cell-based screening.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Suspension Culture, Lymphocyte Of B Lineage, Epithelial Cell, Cell Culture

DISEASE(S): Prostate Adenocarcinoma,Cervix Carcinoma,Lymphoma

SUBMITTER: Daisuke Ogasawara

LAB HEAD: Benjamin F. Cravatt

PROVIDER: PXD050096 | Pride | 2024-05-31

REPOSITORIES: Pride

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	010923_73A_comp_Ramos_22_0503_01.raw	Raw
	010923_73A_comp_Ramos_22_0503_02.raw	Raw
	010923_73A_comp_Ramos_22_0503_03.raw	Raw
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	010923_73A_comp_Ramos_22_0503_05.raw	Raw

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