Abstract
We have reported that peroxynitrite (PON) selectively inactivated prostacyclin synthase (PGIS) by a mechanism of tyrosine nitration at the active site [Zou, Martin and Ullrich (1997) Biol. Chem. Hoppe-Seyler 378, 707-713]. We have now extended our studies on rat mesangial cells (RMC) and show that nitration can occur under the influence of cytokines. Pretreatment of RMC with interleukin 1beta (IL-1beta), which up-regulated cyclo-oxygenase 2 and inducible nitric oxide synthase (NOS-2), significantly attenuated the conversion of [14C]prostaglandin H2 (PGH2) into the stable prostacyclin (PGI2) metabolite 6-oxo-prostaglandin F1alpha (6-oxo-PGF1alpha). The presence of superoxide dismutase (SOD, 100 units/ml) or the NOS synthase inhibitor Nomega-monomethyl-l-arginine (100 microM) as well as cycloheximide (10 microM) plus actinomycin (10 microM) abolished IL-1beta-mediated down-regulation of 6-oxo-PGF1alpha from PGH2. At the same time, 6-oxo-PGF1alpha production from arachidonate (AA) increased at the expense of prostaglandin E2 (PGE2). Neither NO alone generated from different NO donors nor superoxide from xanthine/xanthine oxidase (1-100 m-units/ml) inhibited PGI2 synthesis, either from PGH2 or from AA. Bolus additions of chemically synthesized PON or the PON generator 3-morpholinosydnonimine N-ethylcarbamide (SIN-1) exhibited a potent inhibition of 6-oxo-PGF1alpha release from both PGH2 and AA. In addition, immunoprecipitation of nitrotyrosine-containing proteins from PON- and SIN-1-treated RMC yielded distinct nitrated PGIS bands but also from IL-1beta-pretreated cells alone, compared with a lack of nitrated PGIS in control cells. Taken together, our results strongly suggest that IL-1beta pretreatment of RMC via NOS-2 leads to the production of PON with the consequence of a partial nitration and inhibition of PGIS.
Full Text
The Full Text of this article is available as a PDF (161.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adler S., Baker P. J., Johnson R. J., Ochi R. F., Pritzl P., Couser W. G. Complement membrane attack complex stimulates production of reactive oxygen metabolites by cultured rat mesangial cells. J Clin Invest. 1986 Mar;77(3):762–767. doi: 10.1172/JCI112372. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ardaillou N., Hagege J., Nivez M. P., Ardaillou R., Schlondorff D. Vasoconstrictor-evoked prostaglandin synthesis in cultured human mesangial cells. Am J Physiol. 1985 Feb;248(2 Pt 2):F240–F246. doi: 10.1152/ajprenal.1985.248.2.F240. [DOI] [PubMed] [Google Scholar]
- Baud L., Ardaillou R. Involvement of reactive oxygen species in kidney damage. Br Med Bull. 1993 Jul;49(3):621–629. doi: 10.1093/oxfordjournals.bmb.a072635. [DOI] [PubMed] [Google Scholar]
- Baud L., Hagege J., Sraer J., Rondeau E., Perez J., Ardaillou R. Reactive oxygen production by cultured rat glomerular mesangial cells during phagocytosis is associated with stimulation of lipoxygenase activity. J Exp Med. 1983 Dec 1;158(6):1836–1852. doi: 10.1084/jem.158.6.1836. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beckman J. S., Ischiropoulos H., Zhu L., van der Woerd M., Smith C., Chen J., Harrison J., Martin J. C., Tsai M. Kinetics of superoxide dismutase- and iron-catalyzed nitration of phenolics by peroxynitrite. Arch Biochem Biophys. 1992 Nov 1;298(2):438–445. doi: 10.1016/0003-9861(92)90432-v. [DOI] [PubMed] [Google Scholar]
- Bohn H., Schönafinger K. Oxygen and oxidation promote the release of nitric oxide from sydnonimines. J Cardiovasc Pharmacol. 1989;14 (Suppl 11):S6–12. [PubMed] [Google Scholar]
- 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]
- Granger D. N., Korthuis R. J. Physiologic mechanisms of postischemic tissue injury. Annu Rev Physiol. 1995;57:311–332. doi: 10.1146/annurev.ph.57.030195.001523. [DOI] [PubMed] [Google Scholar]
- Hecker M., Hatzelmann A., Ullrich V. Preparative HPLC purification of prostaglandin endoperoxides and isolation of novel cyclooxygenase-derived arachidonic acid metabolites. Biochem Pharmacol. 1987 Mar 15;36(6):851–855. doi: 10.1016/0006-2952(87)90175-4. [DOI] [PubMed] [Google Scholar]
- Huie R. E., Padmaja S. The reaction of no with superoxide. Free Radic Res Commun. 1993;18(4):195–199. doi: 10.3109/10715769309145868. [DOI] [PubMed] [Google Scholar]
- Klein T., Nüsing R. M., Pfeilschifter J., Ullrich V. Selective inhibition of cyclooxygenase 2. Biochem Pharmacol. 1994 Oct 18;48(8):1605–1610. doi: 10.1016/0006-2952(94)90205-4. [DOI] [PubMed] [Google Scholar]
- Kulmacz R. J., Lands W. E. Stoichiometry and kinetics of the interaction of prostaglandin H synthase with anti-inflammatory agents. J Biol Chem. 1985 Oct 15;260(23):12572–12578. [PubMed] [Google Scholar]
- Landino L. M., Crews B. C., Timmons M. D., Morrow J. D., Marnett L. J. Peroxynitrite, the coupling product of nitric oxide and superoxide, activates prostaglandin biosynthesis. Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15069–15074. doi: 10.1073/pnas.93.26.15069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsunaga K., Oyama T., Nishimura M. Extraction-photometric determination of nitrite in fresh and sea water. Anal Chim Acta. 1972 Jan;58(1):228–230. doi: 10.1016/S0003-2670(00)86875-0. [DOI] [PubMed] [Google Scholar]
- Moncada S., Higgs A. The L-arginine-nitric oxide pathway. N Engl J Med. 1993 Dec 30;329(27):2002–2012. doi: 10.1056/NEJM199312303292706. [DOI] [PubMed] [Google Scholar]
- Moncada S., Vane J. R. Arachidonic acid metabolites and the interactions between platelets and blood-vessel walls. N Engl J Med. 1979 May 17;300(20):1142–1147. doi: 10.1056/NEJM197905173002006. [DOI] [PubMed] [Google Scholar]
- Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
- Needleman P., Turk J., Jakschik B. A., Morrison A. R., Lefkowith J. B. Arachidonic acid metabolism. Annu Rev Biochem. 1986;55:69–102. doi: 10.1146/annurev.bi.55.070186.000441. [DOI] [PubMed] [Google Scholar]
- Pfeilschifter J., Schalkwijk C., Briner V. A., van den Bosch H. Cytokine-stimulated secretion of group II phospholipase A2 by rat mesangial cells. Its contribution to arachidonic acid release and prostaglandin synthesis by cultured rat glomerular cells. J Clin Invest. 1993 Nov;92(5):2516–2523. doi: 10.1172/JCI116860. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pryor W. A., Squadrito G. L. The chemistry of peroxynitrite: a product from the reaction of nitric oxide with superoxide. Am J Physiol. 1995 May;268(5 Pt 1):L699–L722. doi: 10.1152/ajplung.1995.268.5.L699. [DOI] [PubMed] [Google Scholar]
- Radi R., Beckman J. S., Bush K. M., Freeman B. A. Peroxynitrite oxidation of sulfhydryls. The cytotoxic potential of superoxide and nitric oxide. J Biol Chem. 1991 Mar 5;266(7):4244–4250. [PubMed] [Google Scholar]
- Zou M. H., Ullrich V. Peroxynitrite formed by simultaneous generation of nitric oxide and superoxide selectively inhibits bovine aortic prostacyclin synthase. FEBS Lett. 1996 Mar 11;382(1-2):101–104. doi: 10.1016/0014-5793(96)00160-3. [DOI] [PubMed] [Google Scholar]
- Zou M., Martin C., Ullrich V. Tyrosine nitration as a mechanism of selective inactivation of prostacyclin synthase by peroxynitrite. Biol Chem. 1997 Jul;378(7):707–713. doi: 10.1515/bchm.1997.378.7.707. [DOI] [PubMed] [Google Scholar]