Possible mechanism of nitric oxide production from N-hydroxy- -arginine or hydroxylamine by superoxide ion

It has been speculated that N^G-hydroxy- -arginine (OH- -Arg), which is an intermediate in NO production from -arginine, may be converted to NO by superoxide ion. However, there is still no direct evidence for this conversion. In the present study this was investigated using superoxide ion generated either in acellular or cellular systems. It was found that OH- -Arg and hydroxylamine were converted to nitrite and nitrate apparently via NO by superoxide ion in aqueous solution. Arginine remained unaffected. These changes were observed during reaction of chemical substances as well as in a biological system (zymosan-activated macrophages in culture). Superoxide dismutase prevented this transformation. OH- -Arg was also spontaneously hydrolysed to hydroxylamine and -citrulline, however this occurred at pH> 9 only. Activated microsomes (containing different isoforms of cytochrome P450) were unable to replace NO-synthase in its ability to produce OH- -Arg from -arginine. These data support the hypothesis that a pathway alternative to the well-known synthesis of NO by NO-synthase via OH- -Arg exists. This pathway may involve the production of OH- -Arg by NO-synthase and decomposition of OH- -Arg to NO by the action of superoxide ion. Alternatively, hydrolysis of OH- -Arg to hydroxylamine may occur followed by its oxidation to NO, again by superoxide ion.