Generation of peroxynitrite from reaction of N-acetyl-N-nitrosotryptophan with hydrogen peroxide over a wide range of pH values.
The novel reaction of N-acetyl-N-nitrosotryptophan (NANT) with hydrogen peroxide to yield peroxynitrite is demonstrated. Quantum chemical calculations performed at CBS-QB3 level of theory predicted that the reaction of N-nitrosoindole with both H(2)O(2) and its corresponding anion is thermodynamically feasible. At pH 13, the formation of peroxynitrite from the bimolecular reaction of NANT with H(2)O(2) is unequivocally demonstrated by (15)N NMR spectrometry. In order to prove the intermediacy of peroxynitrite from the NANT-H(2)O(2) system at neutral (7.4) and acidic pH (4.5), the characteristic pattern of CIDNP (chemically induced dynamic nuclear polarization) signals were recorded, i.e. enhanced absorption in the (15)N NMR signal of nitrate and emission in the (15)N NMR signal of nitrite. Most interestingly, the NANT-H(2)O(2) system nitrated N-acetyltyrosine at pH 4 via recombination of freely diffusing nitrogen dioxide and tyrosyl radicals, but nitration was negligible at pH 7.4. Since the combination between NANT and H(2)O(2) is slow, endogenous N-nitrosotryptophan residues cannot act as a "carrier" for peroxynitrite.
Kirsch
Michael
M
Institut für Physiologische Chemie, Universitätsklinikum Essen, Hufelandstrasse 55, D-45122 Essen, Germany. michael.kirsch@uni-essen.de
Lehnig
Manfred
M
D006861:Hydrogen Peroxide / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D009602:Nitrosamines / Q000737:chemistry*; D030421:Peroxynitrous Acid / Q000138:chemical synthesis*; D011789:Quantum Theory; D013056:Spectrophotometry, Ultraviolet; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000737:chemistry
C511420:N-acetyl-N-nitrosotryptophan; D009602:Nitrosamines; D030421:Peroxynitrous Acid; D014364:Tryptophan; D006861:Hydrogen Peroxide