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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

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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) [125I]U-II binding, (2) GTPγ[35S] binding, (3) cAMP formation, and (4) intracellular Ca2+. We assess activity of 9 U-II analogues using these paradigms and examine their ability to contract isolated rat aorta. CHOhUT cells bound [125I]U-II with a B max and K d of 1,110±70 fmol/mg protein and 742 pM, respectively. hU-II stimulated GTPγ[35S] binding (pEC50 8.38), optimal at low (0.1 μM) GDP concentrations. The hU-II GTPγ[35S] response was partially PTx sensitive and there was a potent (pEC50 9.23) low efficacy (∼20% inhibition) coupling to adenylyl cyclase. In CHOhUT cells hU-II stimulates calcium release from intracellular stores (pEC50 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 (pEC50 7.67/pKB 7.55) in GTPγ[35S] binding, a full agonist (pEC50 8.11) for increases in intracellular Ca2+ and a competitive antagonist in the rat aorta bioassay (pKB 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.

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Authors and Affiliations

  1. University Department of Cardiovascular Sciences (Pharmacology and Therapeutics Group), Division of Anaesthesia, Critical Care and Pain Management, Leicester Royal Infirmary, Leicester, LE1 5WW, UK

    Wei Song, John McDonald, Jonathan P. Thompson, David J. Rowbotham & David G. Lambert

  2. Department of Experimental and Clinical Medicine, Section of Pharmacology, University of Ferrara, Ferrara, Italy

    Valeria Camarda & Girolamo Calo

  3. Department of Pharmaceutical Sciences and Biotechnology Centre, University of Ferrara, Ferrara, Italy

    Remo Guerrini & Erika Marzola

Authors
  1. Wei Song
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  2. John McDonald
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  3. Valeria Camarda
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  4. Girolamo Calo
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  5. Remo Guerrini
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  6. Erika Marzola
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  7. Jonathan P. Thompson
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  8. David J. Rowbotham
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  9. David G. Lambert
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Corresponding author

Correspondence to David G. Lambert.

Additional information

W. S. and J. M. contributed equally to this work.

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Song, W., McDonald, J., Camarda, V. et al. Cell and tissue responses of a range of Urotensin II analogs at cloned and native urotensin II receptors. Evidence for coupling promiscuity. Naunyn Schmied Arch Pharmacol 373, 148–157 (2006). https://doi.org/10.1007/s00210-006-0057-2

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  • DOI: https://doi.org/10.1007/s00210-006-0057-2

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