Diaphorase can metabolize some vasorelaxants to NO and eliminate NO scavenging effect of 2-phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (PTIO).
Diaphorase was studied as a possible oxidoreductase participating in NO production from some vasorelaxants. In the presence of NADH or NADPH, diaphorase can convert selected NO donors, glycerol trinitrate (GTN) and formaldoxime (FAL) to nitrites and nitrates with NO as an intermediate. This activity of diaphorase was inhibited by diphenyleneiodonium (DPI) (inhibitor of some NADPH-dependent flavoprotein oxidoreductases), while it remained uninhibited by NG-nitro-L-arginine methyl ester (inhibitor of NO synthase) 7-Ethoxyresorufin (inhibitor of cytochrome P-450 1A1 and cytochrome P-450 NADPH-dependent reductase) inhibited the conversion of GTN only. Existence of NO as an intermediate of the reaction was supported by results of electron paramagnetic resonance spectroscopy. In addition to its ability to affect the above mentioned NO donors, diaphorase was able to reduce 2-phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) and thus to eliminate its NO scavenging effect. This activity of diaphorase could also be inhibited by DPI. The reaction of diaphorase with GTN and PTIO was not affected by superoxide dismutase (SOD) or catalase. Reaction of FAL with diaphorase was lowered with SOD by 38 % indicating the partial participation of superoxide anion probably generated by the reaction of diaphorase with NADH or NADPH. Catalase had no effect. Diaphorase could apparently be one of the enzymes participating in the metabolism of studied NO donors to NO. The easy reduction and consequent elimination of PTIO by diaphorase could affect its use as an NO scavenger in biological tissues.
Bartík
P
P
Department of Biochemistry, Faculty of Science, Charles University, Prague, Czech Republic.
Chalupský
K
K
Vavruska
L
L
Muller
B
B
Stoclet
J-C
JC
Entlicher
G
G
D003497:Cyclic N-Oxides / Q000737:chemistry*; D016166:Free Radical Scavengers / Q000737:chemistry*; D007093:Imidazoles / Q000737:chemistry*; D009252:NADPH Dehydrogenase / Q000737:chemistry*; D009569:Nitric Oxide / Q000737:chemistry*; D005996:Nitroglycerin / Q000737:chemistry*; D010091:Oximes / Q000737:chemistry*; D014665:Vasodilator Agents / Q000737:chemistry*
D003497:Cyclic N-Oxides; D016166:Free Radical Scavengers; D007093:Imidazoles; D010091:Oximes; D014665:Vasodilator Agents; C079391:2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide; D009569:Nitric Oxide; C484470:formaldoxime; D009252:NADPH Dehydrogenase; D005996:Nitroglycerin