Peptide nitration by peroxynitrite: characterisation of the nitration sites by liquid chromatography/tandem mass spectrometry

✍ Scribed by Ornella Curcuruto; Luca Rovatti; Antonio M. Pastorino; Mahmoud Hamdan

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 142 KB
• Volume: 13
• Category: Article
• ISSN: 0951-4198
• DOI: 10.1002/(sici)1097-0231(19990215)13:3<156::aid-rcm472>3.0.co;2-m

✦ Synopsis

A number of recent studies have demonstrated that peroxynitrite (ONOO À ) can react with a host of biomolecules, particularly those containing aromatic amino acid residues, resulting in a number of modifications which have been found in association with diverse pathological conditions. Electrospray ionisation tandem mass spectrometry with and without liquid chromatographic separation has been used to examine a series of model peptides following their treatment with peroxynitrite at physiological pH. The mass spectra of sequences containing two tyrosine residues showed the formation of both mononitrated and dinitrated species, while those with phenylalanine showed no detectable nitration. Tryptic digests of two of the investigated peptides were also examined by liquid chromatography/electrospray mass spectrometry, which yielded further information on the competition for NO 2 by multiple nitration sites within a given sequence. In addition, tryptic digests of the mononitrated component of sequences containing tyrosine and tryptophan indicated that mononitration was limited to tyrosine. Furthermore, some of the presented data indicate that, as well as nitrotyrosine and nitrotryptophan formation, the reaction of ONOO À can result in oxidation as well as the formation of labile adducts with NO 2 .