Skip to main content
Log in

UTP is not a biased agonist at human P2Y11 receptors

  • Original Article
  • Published:
Purinergic Signalling Aims and scope Submit manuscript

Abstract

Biased agonism describes a multistate model of G protein-coupled receptor activation in which each ligand induces a unique structural conformation of the receptor, such that the receptor couples differentially to G proteins and other intracellular proteins. P2Y receptors are G protein-coupled receptors that are activated by endogenous nucleotides, such as adenosine 5′-triphosphate (ATP) and uridine 5′-triphosphate (UTP). A previous report suggested that UTP may be a biased agonist at the human P2Y11 receptor, as it increased cytosolic [Ca2+], but did not induce accumulation of inositol phosphates, whereas ATP did both. The mechanism of action of UTP was unclear, so the aim of this study was to characterise the interaction of UTP with the P2Y11 receptor in greater detail. Intracellular Ca2+ was monitored in 1321N1 cells stably expressing human P2Y11 receptors using the Ca2+-sensitive fluorescent indicator, fluo-4. ATP evoked a rapid, concentration-dependent rise in intracellular Ca2+, but surprisingly, even high concentrations of UTP were ineffective. In contrast, UTP was slightly, but significantly more potent than ATP in evoking a rise in intracellular Ca2+ in 1321N1 cells stably expressing the human P2Y2 receptor, with no difference in the maximum response. Thus, the lack of response to UTP at hP2Y11 receptors was not due to a problem with the UTP solution. Furthermore, coapplying a high concentration of UTP with ATP did not inhibit the response to ATP. Thus, contrary to a previous report, we find no evidence for an agonist action of UTP at the human P2Y11 receptor, nor does UTP act as an antagonist.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

1321N1-hP2Y2 cells:

1321N1 cells stably expressing the human P2Y2 receptor

1321N1-hP2Y11 cells:

1321N1 cells stably expressing the human P2Y11 receptor

95 % cl:

95 % confidence limits

ATP:

Adenosine 5′-triphosphate

GPCR:

G protein-coupled receptors

IPs:

Inositol phosphates

UTP:

Uridine 5′-triphosphate

References

  1. Urban JD, Clarke WP, von Zastrow M, Nichols DE, Kobilka B, Weinstein H, Javitch JA, Roth BL, Christopoulos A, Sexton PM, Miller KJ, Spedding M, Mailman RB (2007) Functional selectivity and classical concepts of quantitative pharmacology. J Pharmacol Exp Ther 320:1–13

    Article  CAS  PubMed  Google Scholar 

  2. Gazi L, Nickolls SA, Strange PG (2003) Functional coupling of the human dopamine D2 receptor with Gαi1, Gαi2, Gαi3 and Gαo G proteins: evidence for agonist regulation of G protein selectivity. Br J Pharmacol 138:775–786

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Berg KA, Maayani S, Goldfarb J, Scaramellini C, Leff P, Clarke WP (1998) Effector pathway-dependent relative efficacy at serotonin type 2A and 2C receptors: evidence for agonist-directed trafficking of receptor stimulus. Mol Pharmacol 54:94–104

    CAS  PubMed  Google Scholar 

  4. Liu JJ, Horst R, Katritch V, Stevens RC, Wüthrich K (2012) Biased signaling pathways in β2-adrenergic receptor characterized by 19 F-NMR. Science 335:1106–1110

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Rahmeh R, Damian M, Cottet M, Orcel H, Mendre C, Durroux T, Sharma KS, Durand G, Pucci B, Trinquet E, Zwier JM, Deupi X, Bron P, Banères JL, Mouillac B, Granier S (2012) Structural insights into biased G protein-coupled receptor signaling revealed by fluorescence spectroscopy. Proc Natl Acad Sci U S A 109:6733–6738

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Abbracchio MP, Burnstock G, Boeynaems JM, Barnard EA, Boyer JL, Kennedy C, Miras-Portugal MT, King BF, Gachet C, Jacobson KA, Weisman GA (2005) The recently deorphanized GPR80 (GPR99) proposed to be the P2Y15 receptor is not a genuine P2Y receptor. Trends Pharmacol Sci 26:8–9

    Article  CAS  PubMed  Google Scholar 

  7. Abbracchio MP, Burnstock G, Boeynaems JM, Barnard EA, Boyer JL, Kennedy C, Fumagalli M, Gachet C, Jacobson KA, Weisman GA (2006) International Union of Pharmacology. Update of the P2Y G protein-coupled nucleotide receptors: from molecular mechanisms and pathophysiology to therapy. Pharmacol Rev 58:281–341

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Kennedy C, Chootip K, Mitchell C, Syed NH, Tengah A (2013) P2X and P2Y nucleotide receptors as targets in cardiovascular disease. Future Med Chem 5:431–439

    Article  CAS  PubMed  Google Scholar 

  9. Hoffmann C, Ziegler N, Reiner S, Krasel C, Lohse MJ (2008) Agonist-selective, receptor-specific interaction of human P2Y receptors with β-arrestin-1 and -2. J Biol Chem 283:30933–30941

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  10. White PJ, Webb TE, Boarder MR (2003) Characterization of a Ca2+ response to both UTP and ATP at human P2Y11 receptors: evidence for agonist-specific signalling. Mol Pharmacol 63:1356–1363

    Article  CAS  PubMed  Google Scholar 

  11. Qi A-D, Kennedy C, Harden TK, Nicholas RA (2001) Differential coupling of the human P2Y11 receptor to phospholipase C and adenylyl cyclase. Br J Pharmacol 132:318–326

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Nicholas RA, Watt WC, Lazarowski ER, Li Q, Harden K (1996) Uridine nucleotide selectivity of three phospholipase C-activating P2 receptors: identification of a UDP-selective, a UTP-selective, and an ATP- and UTP-specific receptor. Mol Pharmacol 50:224–229

    CAS  PubMed  Google Scholar 

  13. Lazarowski ER, Watt WC, Stutts JM, Boucher RC, Harden TK (1995) Pharmacological selectivity of the cloned human P2U-purinoceptor: potent activation by diadenosine tetraphosphate. Br J Pharmacol 116:1619–1627

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Communi D, Govaerts C, Parmentier M, Boeynaems J-M (1997) Cloning of a human purinergic P2Y receptor coupled to phospholipase C and adenylyl cyclase. J Biol Chem 272:31969–31973

    Article  CAS  PubMed  Google Scholar 

  15. Communi D, Robaye B, Boeynaems J-M (1999) Pharmacological characterization of the human P2Y11 receptor. Br J Pharmacol 128:1199–1206

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Kennedy C, Herold CL, Qi A, Harden TK, Nicholas RA (2000) ATP, an agonist at the rat P2Y4 receptor, is an antagonist at the human P2Y4 receptor. Mol Pharmacol 57:926–931

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a grant from The Caledonian Research Foundation (CK).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Charles Kennedy.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Morrow, G.B., Nicholas, R.A. & Kennedy, C. UTP is not a biased agonist at human P2Y11 receptors. Purinergic Signalling 10, 581–585 (2014). https://doi.org/10.1007/s11302-014-9418-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11302-014-9418-3

Keywords

Navigation