Characterization of P2X4 receptor agonists and antagonists by calcium influx and radioligand binding studies
| Autor: | Vigneshwaran Namasivayam, Michel Gillard, Ali El-Tayeb, Jürgen Bajorath, Meryem Köse, Sonja Hinz, Aliaa Abdelrahman, Maggi Burton, Anke C. Schiedel, Christa E. Müller, Marc Roger De Ryck |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Purinergic Antagonists Allosteric regulation Pharmacology Biochemistry Cell Line Mice Radioligand Assay 03 medical and health sciences 0302 clinical medicine Calcium flux Radioligand Animals Humans Binding site Receptor Ion Transport Chemistry Rats 030104 developmental biology Purinergic Agonists Calcium Receptors Purinergic P2X4 030217 neurology & neurosurgery Ion channel linked receptors |
| Zdroj: | Biochemical Pharmacology. 125:41-54 |
| ISSN: | 0006-2952 |
| DOI: | 10.1016/j.bcp.2016.11.016 |
| Popis: | Antagonists for ATP-activated P2X4 ion channel receptors are currently in the focus as novel drug targets, in particular for the treatment of neuropathic and inflammatory pain. We stably expressed the human, rat and mouse P2X4 receptors in 1321N1 astrocytoma cells, which is devoid of functional nucleotide receptors, by retroviral transfection, and established monoclonal cell lines. Calcium flux assay conditions were optimized for high-throughput screening resulting in a Z'-factor of >0.8. The application of ready-to-use frozen cells did not negatively affect the results of the calcium assays, which is of great advantage for the screening of compound libraries. Species differences were observed, the rat P2X4 receptor being particularly insensitive to many ATP derivatives. Membrane preparations of the cell lines showed high levels of specific [35S]ATPγS binding with low nonspecific binding ( |
| Databáze: | OpenAIRE |
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