Bright and stable luminescent probes for target engagement profiling in live cells.

Autor: Payne NC; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA.; Department of Chemistry & Chemical Biology, Harvard University, Cambridge, MA, USA., Kalyakina AS; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA.; Karlsruhe Institute of Technology, Institute of Organic Chemistry, Karlsruhe, Germany., Singh K; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA.; Department of Bioengineering, Northeastern University, Boston, MA, USA., Tye MA; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA.; Harvard Graduate School of Arts and Sciences, Cambridge, MA, USA., Mazitschek R; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA. ralph@broad.harvard.edu.; Harvard T.H. Chan School of Public Health, Boston, MA, USA. ralph@broad.harvard.edu.; Broad Institute of MIT and Harvard, Cambridge, MA, USA. ralph@broad.harvard.edu.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2021 Nov; Vol. 17 (11), pp. 1168-1177. Date of Electronic Publication: 2021 Oct 21.
DOI: 10.1038/s41589-021-00877-5
Abstrakt: The pace of progress in biomedical research directly depends on techniques that enable the quantitative interrogation of interactions between proteins and other biopolymers, or with their small-molecule ligands. Time-resolved Förster resonance energy transfer (TR-FRET) assay platforms offer high sensitivity and specificity. However, the paucity of accessible and biocompatible luminescent lanthanide complexes, which are essential reagents for TR-FRET-based approaches, and their poor cellular permeability have limited broader adaptation of TR-FRET beyond homogeneous and extracellular assay applications. Here, we report the development of CoraFluors, a new class of macrotricyclic terbium complexes, which are synthetically readily accessible, stable in biological media and exhibit photophysical and physicochemical properties that are desirable for biological studies. We validate the performance of CoraFluors in cell-free systems, identify cell-permeable analogs and demonstrate their utility in the quantitative domain-selective characterization of Keap1 ligands, as well as in isoform-selective target engagement profiling of HDAC1 inhibitors in live cells.
(© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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