Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1991 Feb 1;273(Pt 3):547–552. doi: 10.1042/bj2730547

N-hydroxylamine is not an intermediate in the conversion of L-arginine to an activator of soluble guanylate cyclase in neuroblastoma N1E-115 cells.

S Pou 1, W S Pou 1, G M Rosen 1, E E el-Fakahany 1
PMCID: PMC1149797  PMID: 1671745

Abstract

This study evaluates the role of N-hydroxylamine (NH2OH) in activating soluble guanylate cyclase in the mouse neuroblastoma clone N1E-115. It has been proposed that NH2OH is a putative intermediate in the biochemical pathway for the generation of nitric oxide (NO)/endothelium-derived relaxing factor (EDRF) from L-arginine. NH2OH caused a time- and concentration-dependent increase in cyclic GMP formation in intact cells. This response was not dependent on Ca2+. In cytosol preparations the activation of guanylate cyclase by L-arginine was dose-dependent and required Ca2+ and NADPH. In contrast, NH2OH itself did not activate cytosolic guanylate cyclase but it inhibited the basal activity of this enzyme in a concentration-dependent manner. The formation of cyclic GMP in the cytosolic fractions in response to NH2OH required the addition of catalase and H2O2. On the other hand, catalase and/or H2O2 lead to a decrease in L-arginine-induced cyclic GMP formation. Furthermore, NH2OH inhibited L-arginine- and sodium nitroprusside-induced cyclic GMP formation in the cytosol. The inhibition of L-arginine-induced cyclic GMP formation in the cytosol by NH2OH was not reversed by the addition of superoxide dismutase. These data strongly suggest that NH2OH is not a putative intermediate in the metabolism of L-arginine to an activator of guanylate cyclase.

Full text

PDF
547

Selected References

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

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Bredt D. S., Snyder S. H. Isolation of nitric oxide synthetase, a calmodulin-requiring enzyme. Proc Natl Acad Sci U S A. 1990 Jan;87(2):682–685. doi: 10.1073/pnas.87.2.682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bredt D. S., Snyder S. H. Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum. Proc Natl Acad Sci U S A. 1989 Nov;86(22):9030–9033. doi: 10.1073/pnas.86.22.9030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Busse R., Trogisch G., Bassenge E. The role of endothelium in the control of vascular tone. Basic Res Cardiol. 1985 Sep-Oct;80(5):475–490. doi: 10.1007/BF01907912. [DOI] [PubMed] [Google Scholar]
  5. Craven P. A., DeRubertis F. R., Pratt D. W. Electron spin resonance study of the role of NO . catalase in the activation of guanylate cyclase by NaN3 and NH2OH. Modulation of enzyme responses by heme proteins and their nitrosyl derivatives. J Biol Chem. 1979 Sep 10;254(17):8213–8222. [PubMed] [Google Scholar]
  6. DeMaster E. G., Raij L., Archer S. L., Weir E. K. Hydroxylamine is a vasorelaxant and a possible intermediate in the oxidative conversion of L-arginine to nitric oxide. Biochem Biophys Res Commun. 1989 Aug 30;163(1):527–533. doi: 10.1016/0006-291x(89)92169-4. [DOI] [PubMed] [Google Scholar]
  7. Deguchi T. Activation of guanylate cyclase in cerebral cortex of rat by hydroxylamine. J Biol Chem. 1977 Jan 25;252(2):596–601. [PubMed] [Google Scholar]
  8. Deguchi T., Saito M., Kono M. Blockade by N-methylhydroxylamine of activation of guanylate cyclase and elevations of guanosine 3',5'-monophosphate levels in nervous tissues. Biochim Biophys Acta. 1978 Nov 15;544(1):8–19. doi: 10.1016/0304-4165(78)90204-0. [DOI] [PubMed] [Google Scholar]
  9. Deguchi T., Yoshioka M. L-Arginine identified as an endogenous activator for soluble guanylate cyclase from neuroblastoma cells. J Biol Chem. 1982 Sep 10;257(17):10147–10151. [PubMed] [Google Scholar]
  10. Furchgott R. F., Zawadzki J. V. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 1980 Nov 27;288(5789):373–376. doi: 10.1038/288373a0. [DOI] [PubMed] [Google Scholar]
  11. Garthwaite J., Charles S. L., Chess-Williams R. Endothelium-derived relaxing factor release on activation of NMDA receptors suggests role as intercellular messenger in the brain. Nature. 1988 Nov 24;336(6197):385–388. doi: 10.1038/336385a0. [DOI] [PubMed] [Google Scholar]
  12. Gorsky L. D., Förstermann U., Ishii K., Murad F. Production of an EDRF-like activity in the cytosol of N1E-115 neuroblastoma cells. FASEB J. 1990 Mar;4(5):1494–1500. doi: 10.1096/fasebj.4.5.2155150. [DOI] [PubMed] [Google Scholar]
  13. Hibbs J. B., Jr, Taintor R. R., Vavrin Z. Macrophage cytotoxicity: role for L-arginine deiminase and imino nitrogen oxidation to nitrite. Science. 1987 Jan 23;235(4787):473–476. doi: 10.1126/science.2432665. [DOI] [PubMed] [Google Scholar]
  14. Hibbs J. B., Jr, Vavrin Z., Taintor R. R. L-arginine is required for expression of the activated macrophage effector mechanism causing selective metabolic inhibition in target cells. J Immunol. 1987 Jan 15;138(2):550–565. [PubMed] [Google Scholar]
  15. Honegger P., Richelson E. Biochemical differentiation of mechanically dissociated mammalian brain in aggregating cell culture. Brain Res. 1976 Jun 11;109(2):335–354. doi: 10.1016/0006-8993(76)90534-5. [DOI] [PubMed] [Google Scholar]
  16. Ignarro L. J., Buga G. M., Wood K. S., Byrns R. E., Chaudhuri G. Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9265–9269. doi: 10.1073/pnas.84.24.9265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ignarro L. J., Byrns R. E., Buga G. M., Wood K. S. Endothelium-derived relaxing factor from pulmonary artery and vein possesses pharmacologic and chemical properties identical to those of nitric oxide radical. Circ Res. 1987 Dec;61(6):866–879. doi: 10.1161/01.res.61.6.866. [DOI] [PubMed] [Google Scholar]
  18. Ignarro L. J. Endothelium-derived nitric oxide: actions and properties. FASEB J. 1989 Jan;3(1):31–36. doi: 10.1096/fasebj.3.1.2642868. [DOI] [PubMed] [Google Scholar]
  19. Ignarro L. J. Endothelium-derived nitric oxide: pharmacology and relationship to the actions of organic nitrate esters. Pharm Res. 1989 Aug;6(8):651–659. doi: 10.1023/a:1015926119947. [DOI] [PubMed] [Google Scholar]
  20. Ignarro L. J., Kadowitz P. J. The pharmacological and physiological role of cyclic GMP in vascular smooth muscle relaxation. Annu Rev Pharmacol Toxicol. 1985;25:171–191. doi: 10.1146/annurev.pa.25.040185.001131. [DOI] [PubMed] [Google Scholar]
  21. Iyengar R., Stuehr D. J., Marletta M. A. Macrophage synthesis of nitrite, nitrate, and N-nitrosamines: precursors and role of the respiratory burst. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6369–6373. doi: 10.1073/pnas.84.18.6369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Katsuki S., Arnold W., Mittal C., Murad F. Stimulation of guanylate cyclase by sodium nitroprusside, nitroglycerin and nitric oxide in various tissue preparations and comparison to the effects of sodium azide and hydroxylamine. J Cyclic Nucleotide Res. 1977 Feb;3(1):23–35. [PubMed] [Google Scholar]
  23. Knowles R. G., Palacios M., Palmer R. M., Moncada S. Formation of nitric oxide from L-arginine in the central nervous system: a transduction mechanism for stimulation of the soluble guanylate cyclase. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5159–5162. doi: 10.1073/pnas.86.13.5159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kwon N. S., Nathan C. F., Gilker C., Griffith O. W., Matthews D. E., Stuehr D. J. L-citrulline production from L-arginine by macrophage nitric oxide synthase. The ureido oxygen derives from dioxygen. J Biol Chem. 1990 Aug 15;265(23):13442–13445. [PubMed] [Google Scholar]
  25. Long C. J., Berkowitz B. A. What is the relationship between the endothelium derived relaxant factor and nitric oxide? Life Sci. 1989;45(1):1–14. doi: 10.1016/0024-3205(89)90429-3. [DOI] [PubMed] [Google Scholar]
  26. Marletta M. A. Mammalian synthesis of nitrite, nitrate, nitric oxide, and N-nitrosating agents. Chem Res Toxicol. 1988 Sep-Oct;1(5):249–257. doi: 10.1021/tx00005a001. [DOI] [PubMed] [Google Scholar]
  27. Marletta M. A., Yoon P. S., Iyengar R., Leaf C. D., Wishnok J. S. Macrophage oxidation of L-arginine to nitrite and nitrate: nitric oxide is an intermediate. Biochemistry. 1988 Nov 29;27(24):8706–8711. doi: 10.1021/bi00424a003. [DOI] [PubMed] [Google Scholar]
  28. Mayer B., Böhme E. Ca2+-dependent formation of an L-arginine-derived activator of soluble guanylyl cyclase in bovine lung. FEBS Lett. 1989 Oct 9;256(1-2):211–214. doi: 10.1016/0014-5793(89)81750-8. [DOI] [PubMed] [Google Scholar]
  29. Mayer B., Schmidt K., Humbert P., Böhme E. Biosynthesis of endothelium-derived relaxing factor: a cytosolic enzyme in porcine aortic endothelial cells Ca2+-dependently converts L-arginine into an activator of soluble guanylyl cyclase. Biochem Biophys Res Commun. 1989 Oct 31;164(2):678–685. doi: 10.1016/0006-291x(89)91513-1. [DOI] [PubMed] [Google Scholar]
  30. McCall T. B., Boughton-Smith N. K., Palmer R. M., Whittle B. J., Moncada S. Synthesis of nitric oxide from L-arginine by neutrophils. Release and interaction with superoxide anion. Biochem J. 1989 Jul 1;261(1):293–296. doi: 10.1042/bj2610293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Murad F., Waldman S., Molina C., Bennett B., Leitman D. Regulation and role of guanylate cyclase-cyclic GMP in vascular relaxation. Prog Clin Biol Res. 1987;249:65–76. [PubMed] [Google Scholar]
  32. Nathan C. F., Stuehr D. J. Does endothelium-derived nitric oxide have a role in cytokine-induced hypotension? J Natl Cancer Inst. 1990 May 2;82(9):726–728. doi: 10.1093/jnci/82.9.726. [DOI] [PubMed] [Google Scholar]
  33. Palacios M., Knowles R. G., Palmer R. M., Moncada S. Nitric oxide from L-arginine stimulates the soluble guanylate cyclase in adrenal glands. Biochem Biophys Res Commun. 1989 Dec 15;165(2):802–809. doi: 10.1016/s0006-291x(89)80037-3. [DOI] [PubMed] [Google Scholar]
  34. Palmer R. M., Ashton D. S., Moncada S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature. 1988 Jun 16;333(6174):664–666. doi: 10.1038/333664a0. [DOI] [PubMed] [Google Scholar]
  35. Palmer R. M., Ferrige A. G., Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature. 1987 Jun 11;327(6122):524–526. doi: 10.1038/327524a0. [DOI] [PubMed] [Google Scholar]
  36. Palmer R. M., Moncada S. A novel citrulline-forming enzyme implicated in the formation of nitric oxide by vascular endothelial cells. Biochem Biophys Res Commun. 1989 Jan 16;158(1):348–352. doi: 10.1016/s0006-291x(89)80219-0. [DOI] [PubMed] [Google Scholar]
  37. Radomski M. W., Palmer R. M., Moncada S. Comparative pharmacology of endothelium-derived relaxing factor, nitric oxide and prostacyclin in platelets. Br J Pharmacol. 1987 Sep;92(1):181–187. doi: 10.1111/j.1476-5381.1987.tb11310.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rapoport R. M., Murad F. Endothelium-dependent and nitrovasodilator-induced relaxation of vascular smooth muscle: role of cyclic GMP. J Cyclic Nucleotide Protein Phosphor Res. 1983;9(4-5):281–296. [PubMed] [Google Scholar]
  39. Rimele T. J., Sturm R. J., Adams L. M., Henry D. E., Heaslip R. J., Weichman B. M., Grimes D. Interaction of neutrophils with vascular smooth muscle: identification of a neutrophil-derived relaxing factor. J Pharmacol Exp Ther. 1988 Apr;245(1):102–111. [PubMed] [Google Scholar]
  40. Salvemini D., de Nucci G., Gryglewski R. J., Vane J. R. Human neutrophils and mononuclear cells inhibit platelet aggregation by releasing a nitric oxide-like factor. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6328–6332. doi: 10.1073/pnas.86.16.6328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schmidt H. H., Seifert R., Böhme E. Formation and release of nitric oxide from human neutrophils and HL-60 cells induced by a chemotactic peptide, platelet activating factor and leukotriene B4. FEBS Lett. 1989 Feb 27;244(2):357–360. doi: 10.1016/0014-5793(89)80562-9. [DOI] [PubMed] [Google Scholar]
  42. Schmidt K., Mayer B., Kukovetz W. R. Effect of calcium on endothelium-derived relaxing factor formation and cGMP levels in endothelial cells. Eur J Pharmacol. 1989 Nov 7;170(3):157–166. doi: 10.1016/0014-2999(89)90536-0. [DOI] [PubMed] [Google Scholar]
  43. Shikano K., Long C. J., Ohlstein E. H., Berkowitz B. A. Comparative pharmacology of endothelium-derived relaxing factor and nitric oxide. J Pharmacol Exp Ther. 1988 Dec;247(3):873–881. [PubMed] [Google Scholar]
  44. Stuehr D. J., Nathan C. F. Nitric oxide. A macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med. 1989 May 1;169(5):1543–1555. doi: 10.1084/jem.169.5.1543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Surichamorn W., Forray C., el-Fakahany E. E. Role of intracellular Ca2+ mobilization in muscarinic and histamine receptor-mediated activation of guanylate cyclase in N1E-115 neuroblastoma cells: assessment of the arachidonic acid release hypothesis. Mol Pharmacol. 1990 Jun;37(6):860–869. [PubMed] [Google Scholar]
  46. Tayeh M. A., Marletta M. A. Macrophage oxidation of L-arginine to nitric oxide, nitrite, and nitrate. Tetrahydrobiopterin is required as a cofactor. J Biol Chem. 1989 Nov 25;264(33):19654–19658. [PubMed] [Google Scholar]
  47. Thomas G., Ramwell P. W. Vascular relaxation mediated by hydroxylamines and oximes: their conversion to nitrites and mechanism of endothelium dependent vascular relaxation. Biochem Biophys Res Commun. 1989 Oct 31;164(2):889–893. doi: 10.1016/0006-291x(89)91542-8. [DOI] [PubMed] [Google Scholar]
  48. Waldman S. A., Murad F. Cyclic GMP synthesis and function. Pharmacol Rev. 1987 Sep;39(3):163–196. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES