This Article Full Text (OnlineFirst PDF) All Versions of this Article: 1087057108326144v1 13/10/986    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by van der Lee, M. M. C. Articles by Zaman, G. J. R. Search for Related Content PubMed PubMed Citation Articles by van der Lee, M. M. C. Articles by Zaman, G. J. R. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB THIOPHENE Social Bookmarking                         What's this?

-Arrestin Recruitment Assay for the Identification of Agonists of the Sphingosine 1-Phosphate Receptor EDG1

^* To whom correspondence should be addressed. E-mail: guido.zaman{at}spcorp.com.

	Abstract

-Arrestin recruitment assays provide a generic assay platform for drug discovery on G-protein-coupled receptors (GPCRs). The PathHunter^TM assay technology developed by DiscoveRx (Fremont, CA) uses enzyme fragment complementation of -galactosidase to measure receptor--arrestin proximity by chemiluminescence. This study describes an agonistic screen on the human endothelial differentiation sphingolipid GPCR 1 (EDG1), also known as S1P1, using PathHunter^TM-arrestin recruitment technology. Screening of a collection of 345,052 compounds yielded 2157 agonistic hits. Only 10 of these compounds showed -arrestin recruitment activity on a nonrelated receptor, indicating high accuracy and specificity of the assay. The authors show that receptor activation with reference agonists can be detected within the same EDG1 PathHunter^TMcell line at the level of -arrestin recruitment, G_i/o protein-mediated inhibition of cyclic adenosine monophosphate (cAMP), and activation of downstream phosphorylation of extracellular signal-regulated protein kinases. The degree of -arrestin recruitment was largely unaffected upon blockade of G_i/o protein signaling with pertussis toxin, whereas kinetic studies demonstrated a lower rate of -arrestin-receptor association. In contrast, inhibition of cAMP and phosphorylation of extracellular signal-regulated protein kinases were fully G_i/o protein regulated. The data indicate that the -arrestin enzyme fragment complementation cell line can be used not only for agonistic screening of GPCRs but also for the identification of "biased ligands" (i.e., compounds that differ in G-protein coupling and -arrestin-mediated cellular effects). (Journal of Biomolecular Screening XXXX:xx-xx)

First published on November 25, 2008, doi:10.1177/1087057108326144

Journal of Biomolecular Screening 2008;13:986.

A more recent version of this article appeared on December 1, 2008


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				L. H.C. Van Lith, J. Oosterom, A. Van Elsas, and G. J.R. Zaman C5a-Stimulated Recruitment of {beta}-Arrestin2 to the Nonsignaling 7-Transmembrane Decoy Receptor C5L2 J Biomol Screen, October 1, 2009; 14(9): 1067 - 1075. [Abstract] [PDF]

				M. M.C. van der Lee, M. Blomenrohr, A. A. van der Doelen, J. W.Y. Wat, N. Smits, B. J. Hanson, C. J. van Koppen, and G. J.R. Zaman Pharmacological Characterization of Receptor Redistribution and {beta}-Arrestin Recruitment Assays for the Cannabinoid Receptor 1 J Biomol Screen, August 1, 2009; 14(7): 811 - 823. [Abstract] [PDF]