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. 1987 Dec 1;105(6):2745–2750. doi: 10.1083/jcb.105.6.2745

Two G-proteins act in series to control stimulus-secretion coupling in mast cells: use of neomycin to distinguish between G-proteins controlling polyphosphoinositide phosphodiesterase and exocytosis

PMCID: PMC2114701  PMID: 2447099

Abstract

Provision of GTP (or other nucleotides capable of acting as ligands for activation of G-proteins) together with Ca2+ (at micromolar concentrations) is both necessary and sufficient to stimulate exocytotic secretion from mast cells permeabilized with streptolysin-O. GTP and its analogues, through their interactions with Gp, also activate polyphosphoinositide-phosphodiesterase (PPI-pde generating inositol 1,4,5-trisphosphate and diglyceride [DG]). We have used mast cells labeled with [3H]inositol to test whether the requirement for GTP in exocytosis is an expression of Gp activity through the generation of DG and consequent activation of protein kinase C, or whether GTP is required at a later stage in the stimulus secretion sequence. Neomycin (0.3 mM) inhibits activation of PPI-pde, but maximal secretion due to optimal concentrations of guanosine 5'-O-(3-thiotriphosphate) (GTP- gamma-S) can still be evoked in its presence. When ATP is also provided the concentration requirement for GTP-gamma-S in support of exocytosis is reduced. This sparing effect of ATP is nullified when the PPI-pde reaction is inhibited by neomycin. We argue that the sparing effect of ATP occurs as a result of enhancement of DG production and through its action as a phosphoryl donor in the reactions catalyzed by protein kinase C.

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Selected References

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  1. Ackermann K. E., Gish B. G., Honchar M. P., Sherman W. R. Evidence that inositol 1-phosphate in brain of lithium-treated rats results mainly from phosphatidylinositol metabolism. Biochem J. 1987 Mar 1;242(2):517–524. doi: 10.1042/bj2420517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrowman M. M., Cockcroft S., Gomperts B. D. Differential control of azurophilic and specific granule exocytosis in Sendai-virus-permeabilized rabbit neutrophils. J Physiol. 1987 Feb;383:115–124. doi: 10.1113/jphysiol.1987.sp016399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barrowman M. M., Cockcroft S., Gomperts B. D. Two roles for guanine nucleotides in the stimulus-secretion sequence of neutrophils. Nature. 1986 Feb 6;319(6053):504–507. doi: 10.1038/319504a0. [DOI] [PubMed] [Google Scholar]
  4. Berridge M. J. Inositol trisphosphate and diacylglycerol as second messengers. Biochem J. 1984 Jun 1;220(2):345–360. doi: 10.1042/bj2200345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bilezikian J. P., Aurbach G. D. The effects of nucleotides on the expression of beta-adrenergic adenylate cyclase activity in membranes from turkey erythrocytes. J Biol Chem. 1974 Jan 10;249(1):157–161. [PubMed] [Google Scholar]
  6. Bittner M. A., Holz R. W., Neubig R. R. Guanine nucleotide effects on catecholamine secretion from digitonin-permeabilized adrenal chromaffin cells. J Biol Chem. 1986 Aug 5;261(22):10182–10188. [PubMed] [Google Scholar]
  7. Cockcroft S., Gomperts B. D. Role of guanine nucleotide binding protein in the activation of polyphosphoinositide phosphodiesterase. Nature. 1985 Apr 11;314(6011):534–536. doi: 10.1038/314534a0. [DOI] [PubMed] [Google Scholar]
  8. Cockcroft S., Taylor J. A., Judah J. D. Subcellular localisation of inositol lipid kinases in rat liver. Biochim Biophys Acta. 1985 May 30;845(2):163–170. doi: 10.1016/0167-4889(85)90173-9. [DOI] [PubMed] [Google Scholar]
  9. Cockcroft S. The dependence on Ca2+ of the guanine-nucleotide-activated polyphosphoinositide phosphodiesterase in neutrophil plasma membranes. Biochem J. 1986 Dec 1;240(2):503–507. doi: 10.1042/bj2400503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Downes C. P., Michell R. H. The polyphosphoinositide phosphodiesterase of erythrocyte membranes. Biochem J. 1981 Jul 15;198(1):133–140. doi: 10.1042/bj1980133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fitzpatrick L. A., Brandi M. L., Aurbach G. D. Calcium-controlled secretion is effected through a guanine nucleotide regulatory protein in parathyroid cells. Endocrinology. 1986 Dec;119(6):2700–2703. doi: 10.1210/endo-119-6-2700. [DOI] [PubMed] [Google Scholar]
  12. Gomperts B. D., Barrowman M. M., Cockcroft S. Dual role for guanine nucleotides in stimulus-secretion coupling. Fed Proc. 1986 Jun;45(7):2156–2161. [PubMed] [Google Scholar]
  13. Gomperts B. D. Involvement of guanine nucleotide-binding protein in the gating of Ca2+ by receptors. Nature. 1983 Nov 3;306(5938):64–66. doi: 10.1038/306064a0. [DOI] [PubMed] [Google Scholar]
  14. Hallcher L. M., Sherman W. R. The effects of lithium ion and other agents on the activity of myo-inositol-1-phosphatase from bovine brain. J Biol Chem. 1980 Nov 25;255(22):10896–10901. [PubMed] [Google Scholar]
  15. Haslam R. J., Davidson M. M. Guanine nucleotides decrease the free [Ca2+] required for secretion of serotonin from permeabilized blood platelets. Evidence of a role for a GTP-binding protein in platelet activation. FEBS Lett. 1984 Aug 20;174(1):90–95. doi: 10.1016/0014-5793(84)81084-4. [DOI] [PubMed] [Google Scholar]
  16. Howell T. W., Cockcroft S., Gomperts B. D. Essential synergy between Ca2+ and guanine nucleotides in exocytotic secretion from permeabilized rat mast cells. J Cell Biol. 1987 Jul;105(1):191–197. doi: 10.1083/jcb.105.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Howell T. W., Gomperts B. D. Rat mast cells permeabilised with streptolysin O secrete histamine in response to Ca2+ at concentrations buffered in the micromolar range. Biochim Biophys Acta. 1987 Feb 18;927(2):177–183. doi: 10.1016/0167-4889(87)90132-7. [DOI] [PubMed] [Google Scholar]
  18. Johnson G. S., Mukku V. R. Evidence in intact cells for an involvement of GTP in the activation of adenylate cyclase. J Biol Chem. 1979 Jan 10;254(1):95–100. [PubMed] [Google Scholar]
  19. Knight D. E., Baker P. F. Guanine nucleotides and Ca-dependent exocytosis. Studies on two adrenal cell preparations. FEBS Lett. 1985 Sep 23;189(2):345–349. doi: 10.1016/0014-5793(85)81053-x. [DOI] [PubMed] [Google Scholar]
  20. Knight D. E., Scrutton M. C. Effect of various excitatory agonists on the secretion of 5-hydroxytryptamine from permeabilised human platelets induced by Ca2+ in the presence or absence of GTP. FEBS Lett. 1985 Apr 22;183(2):417–422. doi: 10.1016/0014-5793(85)80823-1. [DOI] [PubMed] [Google Scholar]
  21. Knight D. E., Scrutton M. C. Effects of guanine nucleotides on the properties of 5-hydroxytryptamine secretion from electropermeabilised human platelets. Eur J Biochem. 1986 Oct 1;160(1):183–190. doi: 10.1111/j.1432-1033.1986.tb09956.x. [DOI] [PubMed] [Google Scholar]
  22. Litosch I., Fain J. N. 5-Methyltryptamine stimulates phospholipase C-mediated breakdown of exogenous phosphoinositides by blowfly salivary gland membranes. J Biol Chem. 1985 Dec 25;260(30):16052–16055. [PubMed] [Google Scholar]
  23. Marquardt D. L., Gruber H. E., Walker L. L. Ribavirin inhibits mast cell mediator release. J Pharmacol Exp Ther. 1987 Jan;240(1):145–149. [PubMed] [Google Scholar]
  24. Martin T. F., Lucas D. O., Bajjalieh S. M., Kowalchyk J. A. Thyrotropin-releasing hormone activates a Ca2+-dependent polyphosphoinositide phosphodiesterase in permeable GH3 cells. GTP gamma S potentiation by a cholera and pertussis toxin-insensitive mechanism. J Biol Chem. 1986 Feb 25;261(6):2918–2927. [PubMed] [Google Scholar]
  25. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  26. Oetting M., LeBoff M., Swiston L., Preston J., Brown E. Guanine nucleotides are potent secretagogues in permeabilized parathyroid cells. FEBS Lett. 1986 Nov 10;208(1):99–104. doi: 10.1016/0014-5793(86)81540-x. [DOI] [PubMed] [Google Scholar]
  27. Schacht J. Purification of polyphosphoinositides by chromatography on immobilized neomycin. J Lipid Res. 1978 Nov;19(8):1063–1067. [PubMed] [Google Scholar]
  28. Streb H., Heslop J. P., Irvine R. F., Schulz I., Berridge M. J. Relationship between secretagogue-induced Ca2+ release and inositol polyphosphate production in permeabilized pancreatic acinar cells. J Biol Chem. 1985 Jun 25;260(12):7309–7315. [PubMed] [Google Scholar]
  29. Uhing R. J., Prpic V., Jiang H., Exton J. H. Hormone-stimulated polyphosphoinositide breakdown in rat liver plasma membranes. Roles of guanine nucleotides and calcium. J Biol Chem. 1986 Feb 15;261(5):2140–2146. [PubMed] [Google Scholar]
  30. Vallar L., Biden T. J., Wollheim C. B. Guanine nucleotides induce Ca2+-independent insulin secretion from permeabilized RINm5F cells. J Biol Chem. 1987 Apr 15;262(11):5049–5056. [PubMed] [Google Scholar]
  31. Wolff J., Cook G. H. Activation of thyroid membrane adenylate cyclase by purine nucleotides. J Biol Chem. 1973 Jan 10;248(1):350–355. [PubMed] [Google Scholar]

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