Nitric oxide formation during light-induced decomposition of phenyl N-tert-butylnitrone.
Phenyl N-tert-butylnitrone (PBN) is a spin trap commonly employed in free radical research. PBN has been shown to have adverse and beneficial effects on various biological systems. We report here evidence that photolysis (or even ambient light) decomposes PBN to nitric oxide in aqueous solutions. Non-heme and heme proteins have been employed to form nitrosyl complexes, which were detected using EPR spectroscopy. Concomitantly, nitrite formation was detected after light-induced decomposition of PBN. In addition, we found that tert-nitrosobutane and decomposed PBN caused an activation of guanylate cyclase. We propose a mechanism where PBN is decomposed by light to tert-nitrosobutane. The latter compound is, in turn, decomposed to nitric oxide. This study suggests the possibility that PBN or PBN radical adducts may be sources of nitric oxide in biological environments. When using PBN as a spin trap in biological samples, not only is the trapping of reactive free radicals operative, but nitric oxide produced from PBN decomposition may play an important role in altering biological functions.
Chamulitrat
W
W
Laboratory of Molecular Biophysics, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709.
Jordan
S J
SJ
Mason
R P
RP
Saito
K
K
Cutler
R G
RG
D000818:Animals; D006152:Cyclic GMP / Q000032:analysis / Q000378:metabolism; D003497:Cyclic N-Oxides; D004578:Electron Spin Resonance Spectroscopy; D004912:Erythrocytes / Q000378:metabolism; D006162:Guanylate Cyclase / Q000378:metabolism*; D006418:Heme / Q000032:analysis; D006420:Hemeproteins / Q000737:chemistry / Q000378:metabolism*; D008027:Light; D008099:Liver / Q000201:enzymology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008861:Microsomes / Q000378:metabolism; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism; D009573:Nitrites / Q000032:analysis*; D009589:Nitrogen Oxides / Q000737:chemistry* / Q000494:pharmacology / Q000528:radiation effects; D009599:Nitroprusside / Q000494:pharmacology; D010782:Photolysis; D011863:Radioimmunoassay; D051381:Rats; D017207:Rats, Sprague-Dawley; D013113:Spin Labels
D003497:Cyclic N-Oxides; D006420:Hemeproteins; D009573:Nitrites; D009589:Nitrogen Oxides; D013113:Spin Labels; D009599:Nitroprusside; D009569:Nitric Oxide; C029217:phenyl-N-tert-butylnitrone; D006418:Heme; D006162:Guanylate Cyclase; D006152:Cyclic GMP