Target engagement and drug residence time can be observed in living cells with BRET.
作者： Matthew B. Robers , Melanie L. Dart , Carolyn C. Woodroofe , Chad A. Zimprich , Thomas A. Kirkland
DOI: 10.1038/NCOMMS10091
关键词:
摘要: The therapeutic action of drugs is predicated on their physical engagement with cellular targets. Here we describe a broadly applicable method using bioluminescence resonance energy transfer (BRET) to reveal the binding characteristics drug selected targets within intact cells. Cell-permeable fluorescent tracers are used in competitive format quantify target proteins fused Nanoluc luciferase. approach enabled us profile isozyme-specific and kinetics for panel histone deacetylase (HDAC) inhibitors. Our analysis was directed particularly clinically approved prodrug FK228 (Istodax/Romidepsin) because its unique largely unexplained mechanism sustained intracellular action. Analysis by BRET revealed remarkably long residence times at HDAC1, explaining protracted behaviour this prodrug. results demonstrate novel application assessing complex milieu environment.
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