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. 1996 Dec;119(8):1533–1540. doi: 10.1111/j.1476-5381.1996.tb16069.x

Regulation of nitric oxide synthesis by dimethylarginine dimethylaminohydrolase.

R J MacAllister 1, H Parry 1, M Kimoto 1, T Ogawa 1, R J Russell 1, H Hodson 1, G S Whitley 1, P Vallance 1
PMCID: PMC1915783  PMID: 8982498

Abstract

1. Dimethylarginine dimethylaminohydrolase (DDAH), an enzyme that metabolizes the endogenous nitric oxide synthase inhibitors NG-monomethyl-arginine and NG,NG-dimethy-L-arginine to citrulline, was identified by Western blotting in rat and human tissue homogenates. 2. S-2-amino-4(3-methylguanidino)butanoic acid (4124W) inhibited the metabolism of [14C]-NG-monomethyl-L-arginine to [14C]-citrulline by rat liver homogenates (IC50 416 +/- 66 microM; n = 9), human cultured endothelial cells (IC50 250 +/- 34 microM; n = 9) and isolated purified dimethylarginine dimethylaminohydrolase. 3. Addition of 4124W to culture medium increased the accumulation of endogenously-generated NG,NG-dimethy-L-arginine in the supernatant of human cultured endothelial cells from 3.1 +/- 0.3 to 5 +/- 0.7 microM (n = 15; P < 0.005). 4. 4124W (1 microM - 1 mM) had no direct effect on endothelial nitric oxide synthase activity but caused endothelium-dependent contraction of rat aortic rings (1 mM 4124W increased tone by 81.5 +/- 9.6% of that caused by phenylephrine 100 nM). This effect was reversed by L-arginine (100 microM). 4124W reversed endothelium-dependent relaxation of human saphenous vein (19.2 +/- 6.7% reversal of bradykinin-induced relaxation at 1 mM 4124W). 5. These data suggest that inhibition of dimethylarginine dimethylaminohydrolase increases the intracellular contraction of NG,NG-dimethyl-L-arginine sufficiently to inhibit nitric oxide synthesis. Inhibiting the activity of DDAH may provide an alternative mechanism for inhibition of nitric oxide synthases and changes in the activity of DDAH could contribute to pathophysiological alterations in NO generation.

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Selected References

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

  1. Aneman A., Backman V., Snygg J., von Bothmer C., Fändriks L., Pettersson A. Accumulation of an endogenous inhibitor of nitric oxide synthase during graded hemorrhagic shock. Circ Shock. 1994 Nov;44(3):111–114. [PubMed] [Google Scholar]
  2. Azuma H., Sato J., Hamasaki H., Sugimoto A., Isotani E., Obayashi S. Accumulation of endogenous inhibitors for nitric oxide synthesis and decreased content of L-arginine in regenerated endothelial cells. Br J Pharmacol. 1995 Jul;115(6):1001–1004. doi: 10.1111/j.1476-5381.1995.tb15910.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Creager M. A., Gallagher S. J., Girerd X. J., Coleman S. M., Dzau V. J., Cooke J. P. L-arginine improves endothelium-dependent vasodilation in hypercholesterolemic humans. J Clin Invest. 1992 Oct;90(4):1248–1253. doi: 10.1172/JCI115987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fickling S. A., Tooze J. A., Whitley G. S. Characterization of human umbilical vein endothelial cell lines produced by transfection with the early region of SV40. Exp Cell Res. 1992 Aug;201(2):517–521. doi: 10.1016/0014-4827(92)90303-p. [DOI] [PubMed] [Google Scholar]
  6. Förstermann U., Closs E. I., Pollock J. S., Nakane M., Schwarz P., Gath I., Kleinert H. Nitric oxide synthase isozymes. Characterization, purification, molecular cloning, and functions. Hypertension. 1994 Jun;23(6 Pt 2):1121–1131. doi: 10.1161/01.hyp.23.6.1121. [DOI] [PubMed] [Google Scholar]
  7. Garthwaite J. Glutamate, nitric oxide and cell-cell signalling in the nervous system. Trends Neurosci. 1991 Feb;14(2):60–67. doi: 10.1016/0166-2236(91)90022-m. [DOI] [PubMed] [Google Scholar]
  8. Garvey E. P., Tuttle J. V., Covington K., Merrill B. M., Wood E. R., Baylis S. A., Charles I. G. Purification and characterization of the constitutive nitric oxide synthase from human placenta. Arch Biochem Biophys. 1994 Jun;311(2):235–241. doi: 10.1006/abbi.1994.1232. [DOI] [PubMed] [Google Scholar]
  9. Kimoto M., Tsuji H., Ogawa T., Sasaoka K. Detection of NG,NG-dimethylarginine dimethylaminohydrolase in the nitric oxide-generating systems of rats using monoclonal antibody. Arch Biochem Biophys. 1993 Feb 1;300(2):657–662. doi: 10.1006/abbi.1993.1091. [DOI] [PubMed] [Google Scholar]
  10. Kimoto M., Whitley G. S., Tsuji H., Ogawa T. Detection of NG,NG-dimethylarginine dimethylaminohydrolase in human tissues using a monoclonal antibody. J Biochem. 1995 Feb;117(2):237–238. doi: 10.1093/jb/117.2.237. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Liew F. Y., Millott S., Parkinson C., Palmer R. M., Moncada S. Macrophage killing of Leishmania parasite in vivo is mediated by nitric oxide from L-arginine. J Immunol. 1990 Jun 15;144(12):4794–4797. [PubMed] [Google Scholar]
  13. MacAllister R. J., Fickling S. A., Whitley G. S., Vallance P. Metabolism of methylarginines by human vasculature; implications for the regulation of nitric oxide synthesis. Br J Pharmacol. 1994 May;112(1):43–48. doi: 10.1111/j.1476-5381.1994.tb13026.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MacAllister R. J., Whitley G. S., Vallance P. Effects of guanidino and uremic compounds on nitric oxide pathways. Kidney Int. 1994 Mar;45(3):737–742. doi: 10.1038/ki.1994.98. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Morris S. M., Jr, Billiar T. R. New insights into the regulation of inducible nitric oxide synthesis. Am J Physiol. 1994 Jun;266(6 Pt 1):E829–E839. doi: 10.1152/ajpendo.1994.266.6.E829. [DOI] [PubMed] [Google Scholar]
  17. Ogawa T., Kimoto M., Sasaoka K. Purification and properties of a new enzyme, NG,NG-dimethylarginine dimethylaminohydrolase, from rat kidney. J Biol Chem. 1989 Jun 15;264(17):10205–10209. [PubMed] [Google Scholar]
  18. Rees D. D., Cellek S., Palmer R. M., Moncada S. Dexamethasone prevents the induction by endotoxin of a nitric oxide synthase and the associated effects on vascular tone: an insight into endotoxin shock. Biochem Biophys Res Commun. 1990 Dec 14;173(2):541–547. doi: 10.1016/s0006-291x(05)80068-3. [DOI] [PubMed] [Google Scholar]
  19. Rees D. D., Palmer R. M., Moncada S. Role of endothelium-derived nitric oxide in the regulation of blood pressure. Proc Natl Acad Sci U S A. 1989 May;86(9):3375–3378. doi: 10.1073/pnas.86.9.3375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ueno S., Sano A., Kotani K., Kondoh K., Kakimoto Y. Distribution of free methylarginines in rat tissues and in the bovine brain. J Neurochem. 1992 Dec;59(6):2012–2016. doi: 10.1111/j.1471-4159.1992.tb10088.x. [DOI] [PubMed] [Google Scholar]
  22. Vallance P., Collier J., Moncada S. Effects of endothelium-derived nitric oxide on peripheral arteriolar tone in man. Lancet. 1989 Oct 28;2(8670):997–1000. doi: 10.1016/s0140-6736(89)91013-1. [DOI] [PubMed] [Google Scholar]
  23. Vallance P., Leone A., Calver A., Collier J., Moncada S. Accumulation of an endogenous inhibitor of nitric oxide synthesis in chronic renal failure. Lancet. 1992 Mar 7;339(8793):572–575. doi: 10.1016/0140-6736(92)90865-z. [DOI] [PubMed] [Google Scholar]
  24. Yu X. J., Li Y. J., Xiong Y. Increase of an endogenous inhibitor of nitric oxide synthesis in serum of high cholesterol fed rabbits. Life Sci. 1994;54(12):753–758. doi: 10.1016/0024-3205(94)00443-9. [DOI] [PubMed] [Google Scholar]

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