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. 1992 Jun 15;284(Pt 3):917–922. doi: 10.1042/bj2840917

G-protein-mediated activation of turkey erythrocyte phospholipase C by beta-adrenergic and P2y-purinergic receptors.

C Vaziri 1, C P Downes 1
PMCID: PMC1132627  PMID: 1352448

Abstract

Isoprenaline, previously known only to stimulate adenylate cyclase via the stimulatory G-protein, Gs, activates turkey erythrocyte ghost phospholipase C (PLC) in a dose-dependent manner when GTP or guanosine 5'-[gamma-thio]triphosphate (GTP[S]) is present. The effect is specific in that it is abolished by beta-adrenergic-receptor antagonists. Stimulation of adenosine receptors, which also couple to adenylate cyclase via Gs in turkey erythrocytes, does not activate PLC, indicating that the stimulation observed in the presence of isoprenaline is not due to Gs activation. Furthermore, the stimulation seen is independent of cyclic AMP production. Purified turkey erythrocyte PLC is activated in an adenosine 5'-[beta-thio]diphosphate (ADP[S]; a P2y-purinergic-receptor agonist)- or isoprenaline-regulated manner when reconstituted with turkey erythrocyte ghosts, demonstrating that a single species of PLC effector enzyme can be regulated by both the purinergic and the beta-adrenergic receptor populations present in turkey erythrocyte membranes. Pretreatment of intact turkey erythrocytes with the P2y agonist ADP[S] causes decreased PLC responsiveness of subsequent ghost preparations to ADP[S] stimulation, although responses to isoprenaline are unaffected (homologous desensitization). In contrast, pretreatment of intact erythrocytes with isoprenaline results in heterologous desensitization of both the P2y and the beta-adrenergic receptors. These effects occur at the level of receptor-G-protein coupling, since PLC stimulation by GTP[S] (which directly activates G-proteins) in the absence of agonists is unaffected.

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Selected References

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  1. Benovic J. L., Strasser R. H., Caron M. G., Lefkowitz R. J. Beta-adrenergic receptor kinase: identification of a novel protein kinase that phosphorylates the agonist-occupied form of the receptor. Proc Natl Acad Sci U S A. 1986 May;83(9):2797–2801. doi: 10.1073/pnas.83.9.2797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boyer J. L., Downes C. P., Harden T. K. Kinetics of activation of phospholipase C by P2Y purinergic receptor agonists and guanine nucleotides. J Biol Chem. 1989 Jan 15;264(2):884–890. [PubMed] [Google Scholar]
  3. Boyer J. L., Waldo G. L., Evans T., Northup J. K., Downes C. P., Harden T. K. Modification of AlF-4- and receptor-stimulated phospholipase C activity by G-protein beta gamma subunits. J Biol Chem. 1989 Aug 15;264(23):13917–13922. [PubMed] [Google Scholar]
  4. Brandt D. R., Ross E. M. Catecholamine-stimulated GTPase cycle. Multiple sites of regulation by beta-adrenergic receptor and Mg2+ studied in reconstituted receptor-Gs vesicles. J Biol Chem. 1986 Feb 5;261(4):1656–1664. [PubMed] [Google Scholar]
  5. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  6. Harden T. K. Agonist-induced desensitization of the beta-adrenergic receptor-linked adenylate cyclase. Pharmacol Rev. 1983 Mar;35(1):5–32. [PubMed] [Google Scholar]
  7. Harden T. K., Hawkins P. T., Stephens L., Boyer J. L., Downes C. P. Phosphoinositide hydrolysis by guanosine 5'-[gamma-thio]triphosphate-activated phospholipase C of turkey erythrocyte membranes. Biochem J. 1988 Jun 1;252(2):583–593. doi: 10.1042/bj2520583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Imai A., Gershengorn M. C. Independent phosphatidylinositol synthesis in pituitary plasma membrane and endoplasmic reticulum. Nature. 1987 Feb 19;325(6106):726–728. doi: 10.1038/325726a0. [DOI] [PubMed] [Google Scholar]
  9. Martin M. W., Harden T. K. Agonist-induced desensitization of a P2Y-purinergic receptor-regulated phospholipase C. J Biol Chem. 1989 Nov 25;264(33):19535–19539. [PubMed] [Google Scholar]
  10. McPhee F., Lowe G., Vaziri C., Downes C. P. Phosphatidylinositol synthase and phosphatidylinositol/inositol exchange reactions in turkey erythrocyte membranes. Biochem J. 1991 Apr 1;275(Pt 1):187–192. doi: 10.1042/bj2750187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Minneman K. P., Weiland G. A., Molinoff P. B. A comparison of the beta-adrenergic receptor of the turkey erythrocyte with mammalian beta1 and beta2 receptors. Mol Pharmacol. 1980 Jan;17(1):1–7. [PubMed] [Google Scholar]
  12. Morris A. J., Waldo G. L., Downes C. P., Harden T. K. A receptor and G-protein-regulated polyphosphoinositide-specific phospholipase C from turkey erythrocytes. I. Purification and properties. J Biol Chem. 1990 Aug 15;265(23):13501–13507. [PubMed] [Google Scholar]
  13. Morris A. J., Waldo G. L., Downes C. P., Harden T. K. A receptor and G-protein-regulated polyphosphoinositide-specific phospholipase C from turkey erythrocytes. II. P2Y-purinergic receptor and G-protein-mediated regulation of the purified enzyme reconstituted with turkey erythrocyte ghosts. J Biol Chem. 1990 Aug 15;265(23):13508–13514. [PubMed] [Google Scholar]
  14. Pike L. J., Lefkowitz R. J. Activation and desensitization of beta-adrenergic receptor-coupled GTPase and adenylate cyclase of frog and turkey erythrocyte membranes. J Biol Chem. 1980 Jul 25;255(14):6860–6867. [PubMed] [Google Scholar]
  15. Rooney T. A., Hager R., Thomas A. P. Beta-adrenergic receptor-mediated phospholipase C activation independent of cAMP formation in turkey erythrocyte membranes. J Biol Chem. 1991 Aug 15;266(23):15068–15074. [PubMed] [Google Scholar]
  16. Smrcka A. V., Hepler J. R., Brown K. O., Sternweis P. C. Regulation of polyphosphoinositide-specific phospholipase C activity by purified Gq. Science. 1991 Feb 15;251(4995):804–807. doi: 10.1126/science.1846707. [DOI] [PubMed] [Google Scholar]
  17. Strathmann M., Simon M. I. G protein diversity: a distinct class of alpha subunits is present in vertebrates and invertebrates. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9113–9117. doi: 10.1073/pnas.87.23.9113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Taylor S. J., Chae H. Z., Rhee S. G., Exton J. H. Activation of the beta 1 isozyme of phospholipase C by alpha subunits of the Gq class of G proteins. Nature. 1991 Apr 11;350(6318):516–518. doi: 10.1038/350516a0. [DOI] [PubMed] [Google Scholar]
  19. Tolkovsky A. M., Levitzki A. Coupling of a single adenylate cyclase to two receptors: adenosine and catecholamine. Biochemistry. 1978 Sep 5;17(18):3811–3817. doi: 10.1021/bi00611a021. [DOI] [PubMed] [Google Scholar]
  20. Waldo G. L., Boyer J. L., Morris A. J., Harden T. K. Purification of an AlF4- and G-protein beta gamma-subunit-regulated phospholipase C-activating protein. J Biol Chem. 1991 Aug 5;266(22):14217–14225. [PubMed] [Google Scholar]

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