| Dokumentenart: | Artikel | ||||||
|---|---|---|---|---|---|---|---|
| Titel eines Journals oder einer Zeitschrift: | Pharmacological Research | ||||||
| Verlag: | ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD | ||||||
| Ort der Veröffentlichung: | LONDON | ||||||
| Band: | 114 | ||||||
| Seitenbereich: | S. 13-26 | ||||||
| Datum: | 11 Oktober 2016 | ||||||
| Zusätzliche Informationen (Öffentlich): | Supporting Information available | ||||||
| Institutionen: | Chemie und Pharmazie > Institut für Pharmazie Chemie und Pharmazie > Institut für Analytische Chemie, Chemo- und Biosensorik > Bioanalytik und Biosensorik (Prof. Joachim Wegener) Chemie und Pharmazie > Institut für Analytische Chemie, Chemo- und Biosensorik > Bioanalytik und Biosensorik (Prof. Joachim Wegener) | ||||||
| Projekte: | GRK 1910, Graduiertenkolleg "Medicinal Chemistry of Selective GPCR Ligands" | ||||||
| Identifikationsnummer: |
| ||||||
| Stichwörter / 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-Dezimal-Klassifikation: | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie 600 Technik, Medizin, angewandte Wissenschaften > 615 Pharmazie | ||||||
| Status: | Veröffentlicht | ||||||
| Begutachtet: | Ja, diese Version wurde begutachtet | ||||||
| An der Universität Regensburg entstanden: | Ja | ||||||
| Dokumenten-ID: | 34702 |
Web of ScienceZusammenfassung
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 ...
Zusammenfassung
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.
Metadaten zuletzt geändert: 28 Mai 2018 06:51
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