Target discovery in small-molecule cell-based screens by in situ proteome reactivity profiling

Michael J. Evans, Alan Saghatelian, Erik J. Sorensen, Benjamin F. Cravatt

Research output: Contribution to journalArticlepeer-review

208 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1303-1307
Number of pages5
JournalNature biotechnology
Volume23
Issue number10
DOIs
StatePublished - Oct 2005

All Science Journal Classification (ASJC) codes

  • Applied Microbiology and Biotechnology
  • Bioengineering
  • Molecular Medicine
  • Biotechnology
  • Biomedical Engineering

Fingerprint

Dive into the research topics of 'Target discovery in small-molecule cell-based screens by in situ proteome reactivity profiling'. Together they form a unique fingerprint.

Cite this