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Cell and tissue responses of a range of Urotensin II analogs at cloned and native urotensin II receptors. Evidence for coupling promiscuity.

Authors :
Song, Wei
McDonald, John
Camarda, Valeria
Calo, Girolamo
Guerrini, Remo
Marzola, Erika
Thompson, Jonathan
Rowbotham, David
Lambert, David
Source :
Naunyn-Schmiedeberg's Archives of Pharmacology; May2006, Vol. 373 Issue 2, p148-157, 10p
Publication Year :
2006

Abstract

Urotensin II (U-II) is the peptide ligand for the G-protein-coupled U-II receptor (UT). U-II has been dubbed “the most potent vasoconstrictor identified to date”. However, in vivo studies with this system are hampered by the paucity of available ligands. Here, we characterise Chinese hamster ovary (CHO) cells expressing the human UT receptor in the following assays; (1) [<superscript>125</superscript>I]U-II binding, (2) GTPγ[<superscript>35</superscript>S] binding, (3) cAMP formation, and (4) intracellular Ca<superscript>2+</superscript>. We assess activity of 9 U-II analogues using these paradigms and examine their ability to contract isolated rat aorta. CHO<subscript>hUT</subscript> cells bound [<superscript>125</superscript>I]U-II with a B <subscript>max</subscript> and K <subscript>d</subscript> of 1,110±70 fmol/mg protein and 742 pM, respectively. hU-II stimulated GTPγ[<superscript>35</superscript>S] binding (pEC<subscript>50</subscript> 8.38), optimal at low (0.1 μM) GDP concentrations. The hU-II GTPγ[<superscript>35</superscript>S] response was partially PTx sensitive and there was a potent (pEC<subscript>50</subscript> 9.23) low efficacy (∼20% inhibition) coupling to adenylyl cyclase. In CHO<subscript>hUT</subscript> cells hU-II stimulates calcium release from intracellular stores (pEC<subscript>50</subscript> 8.80) and calcium influx in a PTx-insensitive manner. In our structure-activity relationship study most ligands acted as full agonists. However, urantide behaved as a partial agonist (pEC<subscript>50</subscript> 7.67/pK<subscript>B</subscript> 7.55) in GTPγ[<superscript>35</superscript>S] binding, a full agonist (pEC<subscript>50</subscript> 8.11) for increases in intracellular Ca<superscript>2+</superscript> and a competitive antagonist in the rat aorta bioassay (pK<subscript>B</subscript> 8.59). Collectively, these data show promiscuity at high expression and indicate the need for careful multi-assay evaluation of novel U-II analogues. Further modification of urantide, in order to eliminate residual agonist activity and to identify novel ligands for in vivo cardiovascular studies are clearly warranted. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
00281298
Volume :
373
Issue :
2
Database :
Complementary Index
Journal :
Naunyn-Schmiedeberg's Archives of Pharmacology
Publication Type :
Academic Journal
Accession number :
20900206
Full Text :
https://doi.org/10.1007/s00210-006-0057-2