TY - JOUR
T1 - Target discovery in small-molecule cell-based screens by in situ proteome reactivity profiling
AU - Evans, Michael J.
AU - Saghatelian, Alan
AU - Sorensen, Erik J.
AU - Cravatt, Benjamin F.
N1 - Funding Information:
We thank J. Tamiya for valuable contributions to the initial design of the probe library and A. Speers for assistance with the identification of the probe labeling site on PGAM1. This work was supported by the National Institutes of Health grant CA087660 (to B.F.C.), the California Breast Cancer Research Foundation (B.F.C.), a Merck Fellowship of the Life Sciences Research Foundation (A.S.) and the Skaggs Institute for Chemical Biology.
PY - 2005/10
Y1 - 2005/10
N2 - Chemical genomics aims to discover small molecules that affect biological processes through the perturbation of protein function. However, determining the protein targets of bioactive compounds remains a formidable challenge. We address this problem here through the creation of a natural product-inspired small-molecule library bearing protein-reactive elements. Cell-based screening identified a compound, MJE3, that inhibits breast cancer cell proliferation. In situ proteome reactivity profiling revealed that MJE3, but not other library members, covalently labeled the glycolytic enzyme phosphoglycerate mutase 1 (PGAM1), resulting in enzyme inhibition. Interestingly, MJE3 labeling and inhibition of PGAM1 were observed exclusively in intact cells. These results support the hypothesis that cancer cells depend on glycolysis for viability and promote PGAM1 as a potential therapeutic target. More generally, the incorporation of protein-reactive compounds into chemical genomics screens offers a means to discover targets of bioactive small molecules in living systems, thereby enabling downstream mechanistic investigations.
AB - Chemical genomics aims to discover small molecules that affect biological processes through the perturbation of protein function. However, determining the protein targets of bioactive compounds remains a formidable challenge. We address this problem here through the creation of a natural product-inspired small-molecule library bearing protein-reactive elements. Cell-based screening identified a compound, MJE3, that inhibits breast cancer cell proliferation. In situ proteome reactivity profiling revealed that MJE3, but not other library members, covalently labeled the glycolytic enzyme phosphoglycerate mutase 1 (PGAM1), resulting in enzyme inhibition. Interestingly, MJE3 labeling and inhibition of PGAM1 were observed exclusively in intact cells. These results support the hypothesis that cancer cells depend on glycolysis for viability and promote PGAM1 as a potential therapeutic target. More generally, the incorporation of protein-reactive compounds into chemical genomics screens offers a means to discover targets of bioactive small molecules in living systems, thereby enabling downstream mechanistic investigations.
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U2 - 10.1038/nbt1149
DO - 10.1038/nbt1149
M3 - Article
C2 - 16200062
AN - SCOPUS:27144510184
SN - 1087-0156
VL - 23
SP - 1303
EP - 1307
JO - Nature biotechnology
JF - Nature biotechnology
IS - 10
ER -