Novel GPCR screening approach: indirect identification of S1P receptor agonists in antagonist screening using a calcium assay.
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Siehler Wagner, Sandra and Guerini, Danilo (2006) Novel GPCR screening approach: indirect identification of S1P receptor agonists in antagonist screening using a calcium assay. Journal of Receptor and Signal Transduction Research, 26 (5-6). pp. 549-575. ISSN 1079-9893
Abstract
To elucidate the physiological function of sphingosine 1-phosphate receptors 1-3 (S1P1-3) we aimed to identify selective ligands for these GPCRs. S1P2 and S1P3 are coupled to Gq, and are, therefore, linked to the phospholipase C/IP3/calcium pathway. S1P1 is solely coupled to Gi and was artificially linked to calcium signaling by coexpression of Galpha 16. The three receptors desensitized on challenge of cells with an agonist (i.e., agonists appeared as antagonists in a second calcium measurement). We screened a compound library for inhibitors of S1P-stimulated calcium signals, and we could identify agonists and antagonists with a single measurement. Agonism and antagonism were confirmed by recording compound-and S1P-induced calcium signals from the same assay well. For the three receptors, we found a reciprocal correlation of agonism and "apparent" antagonism of agonists. In addition, agonists indirectly discovered by this approach do not promote calcium mobilization through endogenous GPCRs.
| Item Type: | Article |
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| Additional Information: | author can archive post-print (ie final draft post-refereeing); 12 month embargo for STM Journals ; 18 month embargo for SSH journals; Publisher's version/PDF cannot be used; Must link to publisher version |
| Keywords: | Gprotein-coupled receptors; Sphingosine1-phosphate receptors; Calcium measurements; Receptor desensitization; Apparent antagonism |
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| Date Deposited: | 14 Dec 2009 13:57 |
| Last Modified: | 31 Jan 2013 01:13 |
| URI: | https://oak.novartis.com/id/eprint/564 |