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Journal of the American Chemical Society DOI:10.1021/ja0352698

Discovery of an inhibitor of a transcription factor using small molecule microarrays and diversity-oriented synthesis.

Publication TypeJournal Article
Year of Publication2003
AuthorsKoehler, AN, Shamji, AF, Schreiber, SL
JournalJournal of the American Chemical Society
Volume125
Issue28
Pages8420-1
Date Published2003/07/16
ISSN0002-7863
Abstract

Small molecule microarrays were screened to identify a small molecule ligand for Hap3p, a subunit of the yeast Hap2/3/4/5p transcription factor complex. The compound, named haptamide A, was determined to have a KD of 5.03 muM for binding to Hap3p using surface plasmon resonance analysis. Haptamide A also inhibited activation of a GDH1-lacZ reporter gene in a dose-dependent fashion. To explore structure-activity relationships, 11 derivatives of haptamide A were prepared using the same synthetic route that was developed for the original library synthesis. Analysis of dissociation constants and IC50 values for the reporter gene assay revealed a more potent inhibitor, haptamide B, with a KD of 330 nM. Whole-genome transcriptional profiling was used to compare effects of haptamide B with a hap3Delta yeast strain. Treatment with haptamide B, like the deletion mutant, reduced lactate-induced transcription of several genes from wild-type levels. Profiling the genetic "knockout" and the chemical genetic "knockdown" led to the identification of several genes that are regulated by Hap3p under nonfermentative conditions. These results demonstrate that a small molecule discovered using the small molecule microarray binding assay can permeate yeast cells and reach its target transcription factor protein in cells.

URLhttp://dx.doi.org/10.1021/ja0352698
DOI10.1021/ja0352698
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/12848532?dopt=Abstract