Graetz, Lukas and Tropmann, Katharina and Bresinsky, Merlin and Mueller, Christoph and Bernhardt, Gunther and Pockes, Steffen (2020) NanoBRET binding assay for histamine H-2 receptor ligands using live recombinant HEK293T cells. SCIENTIFIC REPORTS, 10 (1): 13288. ISSN 2045-2322,
Full text not available from this repository. (Request a copy)Abstract
Fluorescence/luminescence-based techniques play an increasingly important role in the development of test systems for the characterization of future drug candidates, especially in terms of receptor binding in the field of G protein-coupled receptors (GPCRs). In this article, we present the establishment of a homogeneous live cell-based BRET binding assay for the histamine H-2 receptor with different fluorescently labeled squaramide-type compounds synthesized in the course of this study. Py-1-labeled ligand 8 (UR-KAT478) was found to be most suitable in BRET saturation binding experiments with respect to receptor affinity (pK(d) = 7.35) and signal intensity. Real-time kinetic experiments showed a full association of 8 within approximately 30 min and a slow dissociation of the ligand from the receptor. Investigation of reference compounds in BRET-based competition binding with 8 yielded pK(i) values in agreement with radioligand binding data. This study shows that the BRET binding assay is a versatile test system for the characterization of putative new ligands at the histamine H-2 receptor and represents a valuable fluorescence-based alternative to canonical binding assays.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CONSTITUTIVE ACTIVITY; RECEPTOR ANTAGONIST; ACHE INHIBITOR; POTENT; DIZOCILPINE; SELECTIVITY; BRET; |
| Subjects: | 600 Technology > 615 Pharmacy |
| Divisions: | Chemistry and Pharmacy > Institute of Pharmacy Chemistry and Pharmacy > Institute of Pharmacy > Pharmaceutical/Medicinal Chemistry I (Prof. Elz) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 16 Mar 2021 12:22 |
| Last Modified: | 16 Mar 2021 12:22 |
| URI: | https://pred.uni-regensburg.de/id/eprint/44047 |
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