Homogeneous, Real-Time NanoBRET Binding Assays for the Histamine H3 and H4 Receptors on Living Cells.
| Title: | Homogeneous, Real-Time NanoBRET Binding Assays for the Histamine H3 and H4 Receptors on Living Cells. |
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| Authors: | Mocking TAM; Amsterdam Institute for Molecules, Medicines and Systems, Division of Medicinal Chemistry, Faculty of Science, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.; Verweij EWE; Amsterdam Institute for Molecules, Medicines and Systems, Division of Medicinal Chemistry, Faculty of Science, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.; Vischer HF; Amsterdam Institute for Molecules, Medicines and Systems, Division of Medicinal Chemistry, Faculty of Science, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.; Leurs R; Amsterdam Institute for Molecules, Medicines and Systems, Division of Medicinal Chemistry, Faculty of Science, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands r.leurs@vu.nl. |
| Source: | Molecular pharmacology [Mol Pharmacol] 2018 Dec; Vol. 94 (6), pp. 1371-1381. Date of Electronic Publication: 2018 Sep 24. |
| Publication Type: | Journal Article; Research Support, Non-U.S. Gov't |
| Language: | English |
| Journal Info: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 0035623 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1521-0111 (Electronic) Linking ISSN: 0026895X NLM ISO Abbreviation: Mol Pharmacol Subsets: MEDLINE |
| Imprint Name(s): | Publication: 2025- : [Amsterdam] : Elsevier; Original Publication: Bethesda, MD : American Society for Pharmacology and Experimental Therapeutics |
| MeSH Terms: | Biological Assay/*methods ; Protein Binding/*physiology ; Receptors, Histamine H3/*metabolism ; Receptors, Histamine H4/*metabolism; Binding, Competitive/physiology ; Histamine/metabolism ; Radioligand Assay/methods ; Cell Line ; HEK293 Cells ; Humans ; Ligands |
| Abstract: | Receptor-binding affinity and ligand-receptor residence time are key parameters for the selection of drug candidates and are routinely determined using radioligand competition-binding assays. Recently, a novel bioluminescence resonance energy transfer (BRET) method utilizing a NanoLuc-fused receptor was introduced to detect fluorescent ligand binding. Moreover, this NanoBRET method gives the opportunity to follow fluorescent ligand binding on intact cells in real time, and therefore, results might better reflect in vivo conditions as compared with the routinely used cell homogenates or purified membrane fractions. In this study, a real-time NanoBRET-based binding assay was established and validated to detect binding of unlabeled ligands to the histamine H3 receptor (H3R) and histamine H4 receptor on intact cells. Obtained residence times of clinically tested H3R antagonists were reflected by their duration of H3R antagonism in a functional receptor recovery assay.; (Copyright © 2018 by The American Society for Pharmacology and Experimental Therapeutics.) |
| Substance Nomenclature: | 0 (Ligands); 0 (Receptors, Histamine H3); 0 (Receptors, Histamine H4); 820484N8I3 (Histamine) |
| Entry Date(s): | Date Created: 20180926 Date Completed: 20190503 Latest Revision: 20250213 |
| Update Code: | 20260130 |
| DOI: | 10.1124/mol.118.113373 |
| PMID: | 30249614 |
| Database: | MEDLINE |
Journal Article; Research Support, Non-U.S. Gov't