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. 1996 May;118(1):167–173. doi: 10.1111/j.1476-5381.1996.tb15381.x

Second messenger cascade specificity and pharmacological selectivity of the human P2Y1-purinoceptor.

J B Schachter 1, Q Li 1, J L Boyer 1, R A Nicholas 1, T K Harden 1
PMCID: PMC1909474  PMID: 8733591

Abstract

1. The coding sequence of the P2Y1-purinoceptor was cloned from a human genomic library. 2. The open reading frame encodes a protein of 373 amino acids that is 83% identical to the previously cloned chick and turkey P2Y1-purinoceptor and is > or = 95% homologous to the recently cloned rat, mouse, and bovine P2Y1-purinoceptors. 3. The human P2Y1-purinoceptor was stably expressed in 1321N1 human astrocytoma cells using a retroviral vector. Although the P2Y1-purinoceptor agonist, 2MeSATP, had no effect on inositol phosphate accumulation in cells infected with the P2Y1-purinoceptor virus. No effect of 2MeSATP on cyclic AMP accumulation was observed in P2Y1-receptor-expressing 1321N1 cells. 4. The pharmacological selectively of 18 purinoceptor agonists was established for the expressed human P2Y1-purinoceptor. 2MeSATp was more potent than ATP but less potent than 2MeSADP. ADP also was more potent than ATP. A similar maximal effect was observed with most agonists tested. However, alpha, beta-MeATP had no effect and 3'-NH2-3'-deoxyATP and A2P4 were partial agonists. The order of potency of agonists for activation of the turkey P2Y1-purinoceptor, also stably expressed in 1321N1 cells, was identical to that observed for the human P2Y1-purinoceptor. 5. C6 glioma cells express a P2Y-purinoceptor that inhibits adenylyl cyclase but does not activate phospholipase C. Expression of the human P2Y1-purinoceptor in C6 cells conferred 2MeSATP-stimulated inositol lipid hydrolysis to these cells. The phospholipase C-activating human P2Y1-purinoceptor could be delineated from the endogenous P2Y-purinoceptor of C6 glioma cells by use of the P2-purinoceptor antagonist, PPADS, which blocks the P2Y1-purinoceptor but does not block the endogenous P2Y-purinoceptor of C6 cells. P2-purinoceptor agonists also exhibited differential selectivities for activation of these two P2Y-purinoceptors.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abbracchio M. P., Burnstock G. Purinoceptors: are there families of P2X and P2Y purinoceptors? Pharmacol Ther. 1994;64(3):445–475. doi: 10.1016/0163-7258(94)00048-4. [DOI] [PubMed] [Google Scholar]
  2. Boyer J. L., Lazarowski E. R., Chen X. H., Harden T. K. Identification of a P2Y-purinergic receptor that inhibits adenylyl cyclase. J Pharmacol Exp Ther. 1993 Dec;267(3):1140–1146. [PubMed] [Google Scholar]
  3. Boyer J. L., O'Tuel J. W., Fischer B., Jacobson K. A., Harden T. K. Potent agonist action of 2-thioether derivatives of adenine nucleotides at adenylyl cyclase-linked P2Y-purinoceptors. Br J Pharmacol. 1995 Nov;116(6):2611–2616. doi: 10.1111/j.1476-5381.1995.tb17215.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boyer J. L., Zohn I. E., Jacobson K. A., Harden T. K. Differential effects of P2-purinoceptor antagonists on phospholipase C- and adenylyl cyclase-coupled P2Y-purinoceptors. Br J Pharmacol. 1994 Oct;113(2):614–620. doi: 10.1111/j.1476-5381.1994.tb17034.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brake A. J., Wagenbach M. J., Julius D. New structural motif for ligand-gated ion channels defined by an ionotropic ATP receptor. Nature. 1994 Oct 6;371(6497):519–523. doi: 10.1038/371519a0. [DOI] [PubMed] [Google Scholar]
  6. Burnstock G., Kennedy C. Is there a basis for distinguishing two types of P2-purinoceptor? Gen Pharmacol. 1985;16(5):433–440. doi: 10.1016/0306-3623(85)90001-1. [DOI] [PubMed] [Google Scholar]
  7. Chen C. C., Akopian A. N., Sivilotti L., Colquhoun D., Burnstock G., Wood J. N. A P2X purinoceptor expressed by a subset of sensory neurons. Nature. 1995 Oct 5;377(6548):428–431. doi: 10.1038/377428a0. [DOI] [PubMed] [Google Scholar]
  8. Dubyak G. R., el-Moatassim C. Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides. Am J Physiol. 1993 Sep;265(3 Pt 1):C577–C606. doi: 10.1152/ajpcell.1993.265.3.C577. [DOI] [PubMed] [Google Scholar]
  9. Filtz T. M., Li Q., Boyer J. L., Nicholas R. A., Harden T. K. Expression of a cloned P2Y purinergic receptor that couples to phospholipase C. Mol Pharmacol. 1994 Jul;46(1):8–14. [PubMed] [Google Scholar]
  10. Fischer B., Boyer J. L., Hoyle C. H., Ziganshin A. U., Brizzolara A. L., Knight G. E., Zimmet J., Burnstock G., Harden T. K., Jacobson K. A. Identification of potent, selective P2Y-purinoceptor agonists: structure-activity relationships for 2-thioether derivatives of adenosine 5'-triphosphate. J Med Chem. 1993 Nov 26;36(24):3937–3946. doi: 10.1021/jm00076a023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henderson D. J., Elliot D. G., Smith G. M., Webb T. E., Dainty I. A. Cloning and characterisation of a bovine P2Y receptor. Biochem Biophys Res Commun. 1995 Jul 17;212(2):648–656. doi: 10.1006/bbrc.1995.2018. [DOI] [PubMed] [Google Scholar]
  12. Hourani S. M., Cusack N. J. Pharmacological receptors on blood platelets. Pharmacol Rev. 1991 Sep;43(3):243–298. [PubMed] [Google Scholar]
  13. Lazarowski E. R., Harden T. K. Identification of a uridine nucleotide-selective G-protein-linked receptor that activates phospholipase C. J Biol Chem. 1994 Apr 22;269(16):11830–11836. [PubMed] [Google Scholar]
  14. Lazarowski E. R., Watt W. C., Stutts M. J., Boucher R. C., Harden T. K. Pharmacological selectivity of the cloned human P2U-purinoceptor: potent activation by diadenosine tetraphosphate. Br J Pharmacol. 1995 Sep;116(1):1619–1627. doi: 10.1111/j.1476-5381.1995.tb16382.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lustig K. D., Shiau A. K., Brake A. J., Julius D. Expression cloning of an ATP receptor from mouse neuroblastoma cells. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5113–5117. doi: 10.1073/pnas.90.11.5113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Parr C. E., Sullivan D. M., Paradiso A. M., Lazarowski E. R., Burch L. H., Olsen J. C., Erb L., Weisman G. A., Boucher R. C., Turner J. T. Cloning and expression of a human P2U nucleotide receptor, a target for cystic fibrosis pharmacotherapy. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3275–3279. doi: 10.1073/pnas.91.8.3275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tokuyama Y., Hara M., Jones E. M., Fan Z., Bell G. I. Cloning of rat and mouse P2Y purinoceptors. Biochem Biophys Res Commun. 1995 Jun 6;211(1):211–218. doi: 10.1006/bbrc.1995.1798. [DOI] [PubMed] [Google Scholar]
  18. Valera S., Hussy N., Evans R. J., Adami N., North R. A., Surprenant A., Buell G. A new class of ligand-gated ion channel defined by P2x receptor for extracellular ATP. Nature. 1994 Oct 6;371(6497):516–519. doi: 10.1038/371516a0. [DOI] [PubMed] [Google Scholar]
  19. Webb T. E., Simon J., Krishek B. J., Bateson A. N., Smart T. G., King B. F., Burnstock G., Barnard E. A. Cloning and functional expression of a brain G-protein-coupled ATP receptor. FEBS Lett. 1993 Jun 14;324(2):219–225. doi: 10.1016/0014-5793(93)81397-i. [DOI] [PubMed] [Google Scholar]

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