Archives of Pharmacal Research

, Volume 23, Issue 4, pp 407–412 | Cite as

Identification and characterization of nitric oxide synthase inSalmonella typhimurium

  • Don Woong Choi
  • Hye Young Oh
  • Sung Youl Hong
  • Jeung Whan Han
  • Hyang Woo Lee
Research Articles Biomedical Sciences


The presence of the nitric oxide synthase (NOS) enzyme fromSalmonella typhimurium (S. typhimurium) was identified by measuring radiolabeled L-[3H]citrulline and NO, and Western blot analysis. NOS was partially purified by both Mono Q ion exchange and Superose 12HR size exclusion column chromatography, sequentially. The molecular weight of NOS was estimated to be 93.3 kDa by Western blot analysis. The enzyme showed a significant dependency on the typical NOS cofactors; an apparent Km for L-arginine of 34.7 mM and maximum activity between 37°C and 43°C. The activity was inhibited by NOS inhibitors such as aminoguanidine and NG,NG-dimethyl-L-arginine. Taken together, partially purified NOS inS. typhimurium is assumed to be a different isoform of mammalian NOSs.

Key words

Salmonella typhimurium Nitric oxide Nitric oxide synthase 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  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., 72, 248–254 (1976).PubMedCrossRefGoogle Scholar
  2. Bredt, D. S. and Snyder, S. H., Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum.Proc. Natl. Acad. Sci. U.S.A., 86, 9030–9033 (1989).PubMedCrossRefGoogle Scholar
  3. Broman, K., Lauwers, N., Stalon, V. and Wiame, J. M., Oxygen and nitrate in utilization byBacillus licheniformis of the arginase and arginine deiminase routes of arginine catabolism and other factors affecting their synthesis.J. Bacteriol., 135, 920–927 (1978).PubMedGoogle Scholar
  4. Chen, Y. and Rosazza, J. P., A bacterial nitric oxide synthase from aNocardia species.Biochem. Biophys. Res. Commun., 203, 1251–1258 (1994).PubMedCrossRefGoogle Scholar
  5. Chen, Y. and Rosazza, J. P., Purification and characterization of nitric oxide synthase (NOSNoc) from aNocardia species.J. Bacteriol., 177, 5122–5128 (1995).PubMedGoogle Scholar
  6. Choi, W. S., Chang, M. S., Han, J. W., Hong, S. Y. and Lee, H. W., Identification of nitric oxide synthase inStaphylococcus aureus.Biochem. Biophys. Res. Commun., 237, 554–558 (1997).PubMedCrossRefGoogle Scholar
  7. Choi, W. S., Seo, D. W., Chang, M. S., Han, J. W., Hong, S. Y., Paik, W. K. and Lee, H. W., Methylesters of L-arginine and N-nitro-L-arginine induce nitric oxide synthase inStaphylococcus aureus.Biochem. Biophys. Res. Commun., 246, 431–435 (1998).PubMedCrossRefGoogle Scholar
  8. Cohen, J. I., Bartlett, J. A. and Corey, G. R., Extra-intestinal manifestations ofSalmonella infections.Medicine, 66, 349–388 (1987).PubMedCrossRefGoogle Scholar
  9. Hevel, J. M., White, K. A. and Marletta, M. A., Purification of the inducible murine macrophage nitric oxide synthase. Identification as a flavoprotein.J. Biol. Chem., 266, 22789–22791 (1991).PubMedGoogle Scholar
  10. Kalkowski, I. and Conrad, R., Metabolism of nitric oxide in denitrifyingPseudomonas aeruginosa and nitraterespiringBacillus cereus.FEMS Microbiol. Lett., 66, 107–111 (1991).PubMedCrossRefGoogle Scholar
  11. Kiechle, F. L. and Malinski, T., Nitric Oxide. Biochemistry, pathophysiology, and detection.Am. J. Clin. Pathol., 100, 567–575 (1993).PubMedGoogle Scholar
  12. Mayer, B., John, M. and Bohme, E., Purification of a Ca2+/calmodulin-dependent nitric oxide synthase from porcine cerebellum. Cofactor-role of tetrahydrobiopterin.FEBS Lett., 277, 215–219 (1990).PubMedCrossRefGoogle Scholar
  13. Mercenier, A., Stalon, V., Simon, J. P. and Haas, D., Mapping of the arginine deiminase gene inPseudomonas aeruginosa.J. Bacteriol., 149, 787–788 (1982).PubMedGoogle Scholar
  14. Moncada, S., Palmer, R. M. and Higgs, E. A., Nitric oxide: physiology, pathophysiology, and pharmacology.Pharmacol. Rev., 43, 109–142 (1991).PubMedGoogle Scholar
  15. Morita, H., Yoshikawa, H., Sakata, R., Nagata, Y. and Tanaka, H., Synthesis of nitric oxide from the two equivalent guanidino nitrogens of L-arginine byLactobacillus fermentum.J. Bacteriol., 179, 7812–7815 (1997).PubMedGoogle Scholar
  16. Nathan, C., Nitric oxide as a secretory product of mammalian cells.FASEB, J., 6, 3051–3064 (1992).Google Scholar
  17. Palmer, R. M. J., Ashton, D. S. and Moncada, S., Vascular endothelial cells synthesize nitric oxide from L-arginine.Nature 333, 664–666 (1988).PubMedCrossRefGoogle Scholar
  18. Pollock, J. S., Forstermann, U., Mitchell, J. A., Warner, T. D., Schmidt, H. H. H. W., Nakane, M. and Murad, F., Purification and characterization of particulate endothelium-derived relaxing factor synthase from cultured and native bovine aortic endothelial cells.Proc. Natl. Acad. Sci. U.S.A. 88, 10480–10484 (1991).PubMedCrossRefGoogle Scholar
  19. Schmidt, H. H. H. W., Pollock, J. S., Nakane, M., Gorsky, L. D., Forstermann, U. and Murad, F., Purification of a soluble isoform of guanylyl cyclase-activating-factor synthase.Proc. Natl. Acad. Sci. U.S.A., 88, 365–369 (1991).PubMedCrossRefGoogle Scholar
  20. Simon, J. P., Wargnies, B. and Stalon, V., Control of enzyme synthesis in the arginine deiminase pathway ofStreptococcus faecalis.J. Bacteriol., 150, 1085–1090 (1982).PubMedGoogle Scholar
  21. Stuehr, D. J., Cho, H. J., Kwon, N. S., Weise, M. F. and Nathan, C. F., Purification and characterization of the cytokine-induced macrophage nitric oxide synthase: an FAD- and FMN-containing flavoprotein.Proc. Natl. Acad. Sci. U.S.A. 88, 7773–7777 (1991).PubMedCrossRefGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 1999

Authors and Affiliations

  • Don Woong Choi
    • 1
  • Hye Young Oh
    • 2
  • Sung Youl Hong
    • 3
  • Jeung Whan Han
    • 1
  • Hyang Woo Lee
    • 1
  1. 1.College of PharmacySungkyunkwan UniversitySuwon cityKorea
  2. 2.Department of Toxicology, National Institute of Toxicological ResearchKFDASeoulKorea
  3. 3.Department of Genetic Engineering, College of Life Science and Natural ResourcesSungkyunkwan UniversitySuwon cityKorea

Personalised recommendations