Nitric oxide synthase: An endogenous source of elevated nitrite in infected urine

Abstract

To further define the endogenous sources of urine nitrite in urinary tract infections, we measured urinary nitrite levels by the Griess method and assayed urinary nitric oxide (NO) synthase activity by the conversion of 14C-arginine to 14C-citrulline. Endogenous production of 14C-citrulline was confirmed by thin layer chromatography. Exogenous L-arginine increased nitrite production in whole infected urine, but not in bacteria isolated from infected urine. Urinary tract infections significantly increased NO synthase activity in soluble urine fractions, although soluble activity was less than 10% of particulate activity. Urine particulate fractions from women with non-infected urine had greater NO synthase activity than particulate fractions from men with non-infected urine, 11 +/- 2 and 0.2 +/- 0.1 picomol/min/mg protein, respectively. Urinary tract infections increased NO synthase activity in urine particulate fractions from women and men, 99 +/- 20 and 48 +/- 9 picomol/min/mg protein, respectively. The conversion of 14C-arginine to 14C-citrulline required NADPH, was calcium independent, and was inhibited to a greater extent by L-canavanine than by NG-monomethyl-L-arginine or NG-nitro-L-arginine. Human infected urine contains an isoform of NO synthase which is an endogenous source of urine nitrite.