期刊
ACS CHEMICAL BIOLOGY
卷 15, 期 4, 页码 895-903出版社
AMER CHEMICAL SOC
DOI: 10.1021/acschembio.0c00124
关键词
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资金
- National Institutes of Health (NIH) through a NIH Director's Early Independence Award [DP5-OD26380]
ENL is a transcriptional coactivator that recruits elongation machinery to active cis-regulatory elements upon binding of its YEATS domain-a chromatin reader module-to acylated lysine side chains. Discovery chemistry for the ENL YEATS domain is highly motivated by its significance in acute leukemia pathophysiology, but cell-based assays able to support large-scale screening or hit validation efforts do not presently exist. Here, we report on the discovery of a target engagement assay that allows for high-throughput ligand discovery in living cells. This assay is based on the cellular thermal shift assay (CETSA) but does not require exposing cells to elevated temperatures, as small-molecule ligands are able to stabilize the ENL YEATS domain at 37 degrees C. By eliminating temperature shifts, we developed a simplified target engagement assay that requires just two steps: drug treatment and luminescence detection. To demonstrate its value for higher throughput applications, we miniaturized the assay to a 1536-well format and screened 37 120 small molecules, ultimately identifying an aryl-lysine-competitive ENL/AF9 YEATS domain inhibitor.
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