| Item type: | Article | ||||||
|---|---|---|---|---|---|---|---|
| Journal or Publication Title: | Pharmacological Research | ||||||
| Publisher: | ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD | ||||||
| Place of Publication: | LONDON | ||||||
| Volume: | 114 | ||||||
| Page Range: | pp. 13-26 | ||||||
| Date: | 11 October 2016 | ||||||
| Additional Information (public): | Supporting Information available | ||||||
| Institutions: | Chemistry and Pharmacy > Institute of Pharmacy Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Bioanalytik und Biosensorik (Prof. Joachim Wegener) Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Bioanalytik und Biosensorik (Prof. Joachim Wegener) | ||||||
| Projects: | GRK 1910, Graduiertenkolleg "Medicinal Chemistry of Selective GPCR Ligands" | ||||||
| Identification Number: |
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| Keywords: | PROTEIN-COUPLED RECEPTORS; BETA-GAMMA-SUBUNIT; LIVING CELLS; H-1-RECEPTOR AGONISTS; PERTUSSIS TOXIN; GENE-EXPRESSION; GPCR ASSAYS; GUINEA-PIG; SELECTIVITY; ACTIVATION; Histamine H-1 receptor; Impedimetry; Dynamic mass redistribution; Calcium assay; beta-Arrestin recruitment; Reporter gene assay | ||||||
| Dewey Decimal Classification: | 500 Science > 540 Chemistry & allied sciences 500 Science > 570 Life sciences 600 Technology > 615 Pharmacy | ||||||
| Status: | Published | ||||||
| Refereed: | Yes, this version has been refereed | ||||||
| Created at the University of Regensburg: | Yes | ||||||
| Item ID: | 34702 |
Web of ScienceAbstract
A set of histamine H-1 receptor (H-1 R) agonists and antagonists was characterized in functional assays, using dynamic mass redistribution (DMR), electric cell-substrate impedance sensing (ECIS) and various signaling pathway specific readouts (Fura-2 and aequorin calcium assays, arrestin recruitment (luciferase fragment complementation) assay, luciferase gene reporter assay). Data were gained ...
Abstract
A set of histamine H-1 receptor (H-1 R) agonists and antagonists was characterized in functional assays, using dynamic mass redistribution (DMR), electric cell-substrate impedance sensing (ECIS) and various signaling pathway specific readouts (Fura-2 and aequorin calcium assays, arrestin recruitment (luciferase fragment complementation) assay, luciferase gene reporter assay). Data were gained from genetically engineered HEK293T cells and compared with reference data from GTPase assays and radioligand binding. Histamine and the other H-1 R agonists gave different assay-related pEC(50) values, however, the order of potency was maintained. In the luciferase fragment complementation assay, the H1R preferred beta-arrestin2 over beta-arrestin1. The calcium and the impedimetric assay depended on G(q) coupling of the H1R, as demonstrated by complete inhibition of the histamine-induced signals in the presence of the Gq inhibitor FR900359 (UBO-QIC). Whereas partial inhibition by FR900359 was observed in DMR and the gene reporter assay, pertussis toxin substantially decreased the response in DMR, but increased the luciferase signal, reflecting the contribution of both, Gq and Gi, to signaling in these assays. For antagonists, the results from DMR were essentially compatible with those from conventional readouts, whereas the impedance-based data revealed a trend towards higher pK(b) values. ECIS and calcium assays apparently only reflect Gq signaling, whereas DMR and gene reporter assays appear to integrate both, G(q) and G(i) mediated signaling. The results confirm the value of the label-free methods, DMR and ECIS, for the characterization of H1R ligands. Both noninvasive techniques are complementary to each other, but cannot fully replace reductionist signaling pathway focused assays. (C) 2016 Elsevier Ltd. All rights reserved.
Metadata last modified: 28 May 2018 06:51
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