Chemical tools to expand the ligandable proteome: Diversity-oriented synthesis-based photoreactive stereoprobes

TY - JOUR

T1 - Chemical tools to expand the ligandable proteome

T2 - Diversity-oriented synthesis-based photoreactive stereoprobes

AU - Ogasawara, Daisuke

AU - Konrad, David B.

AU - Tan, Zher Yin

AU - Carey, Kimberly L.

AU - Luo, Jessica

AU - Won, Sang Joon

AU - Li, Haoxin

AU - Carter, Trever R.

AU - DeMeester, Kristen E.

AU - Njomen, Evert

AU - Schreiber, Stuart L.

AU - Xavier, Ramnik J.

AU - Melillo, Bruno

AU - Cravatt, Benjamin F.

N1 - Publisher Copyright: © 2024 Elsevier Ltd

PY - 2024/12/19

Y1 - 2024/12/19

N2 - Chemical proteomics enables the global analysis 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, remained 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 screening and chemical proteomics identified photo-stereoprobes that modulate autophagy by engaging the mitochondrial serine protease CLPP. Our findings show the utility of DOS-inspired photo-stereoprobes for expanding the ligandable proteome, furnishing target engagement assays, and facilitating the discovery and characterization of bioactive compounds in phenotypic screens.

AB - Chemical proteomics enables the global analysis 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, remained 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 screening and chemical proteomics identified photo-stereoprobes that modulate autophagy by engaging the mitochondrial serine protease CLPP. Our findings show the utility of DOS-inspired photo-stereoprobes for expanding the ligandable proteome, furnishing target engagement assays, and facilitating the discovery and characterization of bioactive compounds in phenotypic screens.

KW - autophagy

KW - chemical proteomics

KW - diazirine

KW - diversity-oriented synthesis

KW - ligands

KW - NanoBRET

KW - phenotypic screening

KW - photoreactive

KW - probes

KW - proteomics

KW - stereochemistry

UR - https://www.scopus.com/pages/publications/85210768100

U2 - 10.1016/j.chembiol.2024.10.005

DO - 10.1016/j.chembiol.2024.10.005

M3 - Article

C2 - 39547236

AN - SCOPUS:85210768100

SN - 2451-9456

VL - 31

SP - 2138-2155.e32

JO - Cell Chemical Biology

JF - Cell Chemical Biology

IS - 12

ER -