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. 1993 Nov 15;296(Pt 1):169–174. doi: 10.1042/bj2960169

Stimulation of secretion in permeabilized PC12 cells by adenosine 5'-[gamma-thio]triphosphate: possible involvement of nucleoside diphosphate kinase.

N D Vu 1, P D Wagner 1
PMCID: PMC1137670  PMID: 8250839

Abstract

The addition of Ca2+, adenosine 5'-[gamma-thio]triphosphate (ATP[S]) or guanosine 5'-[gamma-thio]triphosphate (GTP[S]) to digitonin-permeabilized PC12 cells stimulates noradrenaline secretion. Both ATP[S] and GTP[S] stimulate release in the absence of Ca2+. Whereas ADP and adenosine 5'-[beta gamma-imido]triphosphate inhibited ATP[S]-stimulated release, they did not inhibit Ca(2+)-stimulated release even in the absence of added ATP. This suggests that the kinase which uses ATP[S] to induce secretion may not play an essential role in Ca(2+)-stimulated release. As GTP[S]-stimulated and ATP[S]-stimulated secretions were not additive, it seemed possible that stimulation by ATP[S] might in part result from the thiophosphorylation of GDP by nucleoside-diphosphate (NDP) kinase to form GTP[S]. The following results are consistent with this possibility. (1) A low concentration of GDP increased ATP[S]-stimulated secretion, but not GTP[S]-stimulated or Ca(2+)-stimulated secretion. (2) A variety of ribo- and deoxyribo-nucleoside di- and tri-phosphates inhibited ATP[S]-stimulated secretion, but not GTP[S]-stimulated or Ca(2+)-stimulated secretion. Thus, like NDP kinase, the kinase which uses ATP[S] to cause noradrenaline release appears to have a very low specificity for ATP. (3) Incubation of permeabilized cells in a sucrose-containing buffer resulted in the preferential loss of ATP[S]-stimulated secretion and a decrease in the level of NDP kinase. The addition of rat liver NDP kinase to those depleted cells partially restored ATP[S]-stimulated secretion.

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Selected References

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  1. Carroll A. G., Rhoads A. R., Wagner P. D. Hydrolysis-resistant GTP analogs stimulate catecholamine release from digitonin-permeabilized PC12 cells. J Neurochem. 1990 Sep;55(3):930–936. doi: 10.1111/j.1471-4159.1990.tb04580.x. [DOI] [PubMed] [Google Scholar]
  2. Edelman A. M., Blumenthal D. K., Krebs E. G. Protein serine/threonine kinases. Annu Rev Biochem. 1987;56:567–613. doi: 10.1146/annurev.bi.56.070187.003031. [DOI] [PubMed] [Google Scholar]
  3. Fabiato A., Fabiato F. Calculator programs for computing the composition of the solutions containing multiple metals and ligands used for experiments in skinned muscle cells. J Physiol (Paris) 1979;75(5):463–505. [PubMed] [Google Scholar]
  4. Gomperts B. D. GE: a GTP-binding protein mediating exocytosis. Annu Rev Physiol. 1990;52:591–606. doi: 10.1146/annurev.ph.52.030190.003111. [DOI] [PubMed] [Google Scholar]
  5. Greene L. A., Rein G. Release, storage and uptake of catecholamines by a clonal cell line of nerve growth factor (NGF) responsive pheo-chromocytoma cells. Brain Res. 1977 Jul 1;129(2):247–263. doi: 10.1016/0006-8993(77)90005-1. [DOI] [PubMed] [Google Scholar]
  6. Hay J. C., Martin T. F. Resolution of regulated secretion into sequential MgATP-dependent and calcium-dependent stages mediated by distinct cytosolic proteins. J Cell Biol. 1992 Oct;119(1):139–151. doi: 10.1083/jcb.119.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Inoue K., Nakazawa K., Fujimori K., Takanaka A. Extracellular adenosine 5'-triphosphate-evoked norepinephrine secretion not relating to voltage-gated Ca channels in pheochromocytoma PC12 cells. Neurosci Lett. 1989 Dec 4;106(3):294–299. doi: 10.1016/0304-3940(89)90179-1. [DOI] [PubMed] [Google Scholar]
  8. Jakobs K. H., Wieland T. Evidence for receptor-regulated phosphotransfer reactions involved in activation of the adenylate cyclase inhibitory G protein in human platelet membranes. Eur J Biochem. 1989 Jul 15;183(1):115–121. doi: 10.1111/j.1432-1033.1989.tb14903.x. [DOI] [PubMed] [Google Scholar]
  9. Kikkawa S., Takahashi K., Takahashi K., Shimada N., Ui M., Kimura N., Katada T. Conversion of GDP into GTP by nucleoside diphosphate kinase on the GTP-binding proteins. J Biol Chem. 1990 Dec 15;265(35):21536–21540. [PubMed] [Google Scholar]
  10. Kimura N., Shimada N. Direct interaction between membrane-associated nucleoside diphosphate kinase and GTP-binding protein(Gs), and its regulation by hormones and guanine nucleotides. Biochem Biophys Res Commun. 1988 Feb 29;151(1):248–256. doi: 10.1016/0006-291x(88)90586-4. [DOI] [PubMed] [Google Scholar]
  11. Kimura N., Shimada N. Evidence for complex formation between GTP binding protein(Gs) and membrane-associated nucleoside diphosphate kinase. Biochem Biophys Res Commun. 1990 Apr 16;168(1):99–106. doi: 10.1016/0006-291x(90)91680-q. [DOI] [PubMed] [Google Scholar]
  12. Kimura N., Shimada N. GDP does not mediate but rather inhibits hormonal signal to adenylate cyclase. J Biol Chem. 1983 Feb 25;258(4):2278–2283. [PubMed] [Google Scholar]
  13. Kimura N., Shimada N. Membrane-associated nucleoside diphosphate kinase from rat liver. Purification, characterization, and comparison with cytosolic enzyme. J Biol Chem. 1988 Apr 5;263(10):4647–4653. [PubMed] [Google Scholar]
  14. Kimura N., Shimada N., Tsubokura M. Adenosine 5'-(beta, gamma-imino)triphosphate and guanosine 5'-O-(2-thiodiphosphate) do not necessarily provide non-phosphorylating conditions in adenylate cyclase studies. Biochem Biophys Res Commun. 1985 Feb 15;126(3):983–991. doi: 10.1016/0006-291x(85)90282-7. [DOI] [PubMed] [Google Scholar]
  15. Matthies H. J., Palfrey H. C., Miller R. J. Calmodulin- and protein phosphorylation-independent release of catecholamines from PC-12 cells. FEBS Lett. 1988 Mar 14;229(2):238–242. doi: 10.1016/0014-5793(88)81132-3. [DOI] [PubMed] [Google Scholar]
  16. Meldolesi J., Huttner W. B., Tsien R. Y., Pozzan T. Free cytoplasmic Ca2+ and neurotransmitter release: studies on PC12 cells and synaptosomes exposed to alpha-latrotoxin. Proc Natl Acad Sci U S A. 1984 Jan;81(2):620–624. doi: 10.1073/pnas.81.2.620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ohtsuki K., Ikeuchi T., Yokoyama M. Characterization of nucleoside-diphosphate kinase-associated guanine nucleotide-binding proteins from HeLa S3 cells. Biochim Biophys Acta. 1986 Jul 16;882(3):322–330. doi: 10.1016/0304-4165(86)90254-0. [DOI] [PubMed] [Google Scholar]
  18. Ohtsuki K., Yokoyama M., Uesaka H. Physiological correlation between nucleoside-diphosphate kinases and the 21-kDa guanine-nucleotide binding proteins copurified with the enzymes from the cell membrane fractions of Ehrlich ascites tumor cells. Biochim Biophys Acta. 1987 Jul 29;929(3):231–238. doi: 10.1016/0167-4889(87)90248-5. [DOI] [PubMed] [Google Scholar]
  19. Otero A. S., Breitwieser G. E., Szabo G. Activation of muscarinic potassium currents by ATP gamma S in atrial cells. Science. 1988 Oct 21;242(4877):443–445. doi: 10.1126/science.3051383. [DOI] [PubMed] [Google Scholar]
  20. Peppers S. C., Holz R. W. Catecholamine secretion from digitonin-treated PC12 cells. Effects of Ca2+, ATP, and protein kinase C activators. J Biol Chem. 1986 Nov 5;261(31):14665–14669. [PubMed] [Google Scholar]
  21. Schäfer T., Karli U. O., Gratwohl E. K., Schweizer F. E., Burger M. M. Digitonin-permeabilized cells are exocytosis competent. J Neurochem. 1987 Dec;49(6):1697–1707. doi: 10.1111/j.1471-4159.1987.tb02427.x. [DOI] [PubMed] [Google Scholar]
  22. Seifert R., Rosenthal W., Schultz G., Wieland T., Gierschick P., Jakobs K. H. The role of nucleoside-diphosphate kinase reactions in G protein activation of NADPH oxidase by guanine and adenine nucleotides. Eur J Biochem. 1988 Jul 15;175(1):51–55. doi: 10.1111/j.1432-1033.1988.tb14165.x. [DOI] [PubMed] [Google Scholar]
  23. Sela D., Ram E., Atlas D. ATP receptor. A putative receptor-operated channel in PC-12 cells. J Biol Chem. 1991 Sep 25;266(27):17990–17994. [PubMed] [Google Scholar]
  24. Wagner P. D., Vu N. D. Regulation of norepinephrine secretion in permeabilized PC12 cells by Ca2(+)-stimulated phosphorylation. Effects of protein phosphatases and phosphatase inhibitors. J Biol Chem. 1990 Jun 25;265(18):10352–10357. [PubMed] [Google Scholar]
  25. Wagner P. D., Vu N. D. Thiophosphorylation causes Ca2+-independent norepinephrine secretion from permeabilized PC12 cells. J Biol Chem. 1989 Nov 25;264(33):19614–19620. [PubMed] [Google Scholar]
  26. Wieland T., Jakobs K. H. Receptor-regulated formation of GTP[gamma S] with subsequent persistent Gs-protein activation in membranes of human platelets. FEBS Lett. 1989 Mar 13;245(1-2):189–193. doi: 10.1016/0014-5793(89)80219-4. [DOI] [PubMed] [Google Scholar]
  27. Yang Y. C., Vu N. D., Wagner P. D. Guanine nucleotide stimulation of norepinephrine secretion from permeabilized PC12 cells: effects of Mg2+, other nucleotide triphosphates and N-ethylmaleimide. Biochim Biophys Acta. 1992 Apr 7;1134(3):285–291. doi: 10.1016/0167-4889(92)90188-h. [DOI] [PubMed] [Google Scholar]

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