Advances in BRET probes for intracellular target engagement studies.
| Title: | Advances in BRET probes for intracellular target engagement studies. |
|---|---|
| Authors: | Capener JL; Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.; Schwalm MP; Promega, Madison, WI, USA.; Vasta JD; Promega, Madison, WI, USA.; Michaud A; Promega, Madison, WI, USA.; Teske KA; Promega, Madison, WI, USA.; Marsiglia WM; University of Alabama at Birmingham, Birmingham, AL, USA.; Huber KVM; Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, UK.; Dar AC; Program in Chemical Biology, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, USA.; Knapp S; Institute for Pharmaceutical Chemistry, Johann Wolfgang Goethe University, Frankfurt am Main, Germany. knapp@pharmchem.uni-frankfurt.de.; Structural Genomics Consortium (SGC), Buchmann Institute for Life Sciences, Frankfurt am Main, Germany. knapp@pharmchem.uni-frankfurt.de.; German Cancer Consortium (DKTK)/German Cancer Research Center (DKFZ), DTKT Site Frankfurt-Mainz, Heidelberg, Germany. knapp@pharmchem.uni-frankfurt.de.; Axtman AD; Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. alison.axtman@unc.edu.; Robers MB; Promega, Madison, WI, USA. matt.robers@promega.com. |
| Source: | Nature chemical biology [Nat Chem Biol] 2026 May; Vol. 22 (5), pp. 691-701. Date of Electronic Publication: 2026 Jan 06. |
| Publication Type: | Journal Article; Review |
| Language: | English |
| Journal Info: | Publisher: Nature Pub. Group Country of Publication: United States NLM ID: 101231976 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1552-4469 (Electronic) Linking ISSN: 15524450 NLM ISO Abbreviation: Nat Chem Biol Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: New York, NY : Nature Pub. Group, [2005]- |
| MeSH Terms: | Bioluminescence Resonance Energy Transfer Techniques*/methods ; Molecular Probes*/chemistry; Drug Discovery/methods ; Small Molecule Libraries/chemistry ; Small Molecule Libraries/pharmacology ; Humans ; Animals ; Protein Binding |
| Abstract: | Assessing drug-target engagement in living cells is essential for verifying the activity of pharmacological agents. Traditional binding assays often overlook key factors such as permeability, intracellular distribution and complex formation that influence target occupancy in cells. Bioluminescence resonance energy transfer (BRET)-based probes enable direct, quantitative assessment of small-molecule binding to proteins in live, intact cells. BRET provides sensitive detection of target engagement across a wide range of target classes, including less-tractable proteins in membrane compartments. Compared to other existing methods, BRET offers quantification of drug occupancy at steady state and open system regimens. Recent innovations in this platform have expanded its utility beyond occupancy confirmation to include applications in polypharmacology and mechanism-of-action studies. Here, we provide an updated perspective on BRET target engagement assays as versatile tools for chemical biology and early-stage drug discovery.; (© 2026. Springer Nature America, Inc.) |
| Competing Interests: | Competing interests: J.D.V., A.M., K.A.T. and M.B.R. are employees of Promega, which owns patents related to NanoBRET. |
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| Grant Information: | 100001797 Pharmaceutical Research and Manufacturers of America Foundation (PhRMA Foundation); P30 CA008748 United States CA NCI NIH HHS; R01 CA258736 United States CA NCI NIH HHS; R01 CA227636 United States CA NCI NIH HHS; R01 CA256480 United States CA NCI NIH HHS |
| Substance Nomenclature: | 0 (Molecular Probes); 0 (Small Molecule Libraries) |
| Entry Date(s): | Date Created: 20260106 Date Completed: 20260429 Latest Revision: 20260429 |
| Update Code: | 20260430 |
| PubMed Central ID: | PMC13045654 |
| DOI: | 10.1038/s41589-025-02103-y |
| PMID: | 41495225 |
| Database: | MEDLINE |
Journal Article; Review