Premium
Monitoring nitrotyrosinylation of a synthetic peptide by capillary zone electrophoresis
Author(s) -
Bakhøj Annette,
Heegaard Niels H. H.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990801)20:12<2519::aid-elps2519>3.0.co;2-4
Subject(s) - capillary electrophoresis , chemistry , tetranitromethane , peptide , chromatography , reagent , pentapeptide repeat , gel electrophoresis , tyrosine , biochemistry , organic chemistry
Proteins may be nitrated on tyrosyl residues (nitrotyrosinylated) by the action of reactive nitrogen species in inflamed tissues. Capillary electrophoresis was used to monitor this reaction in a model system with tetranitromethane as the nitrotyrosinylating reagent and a synthetic pentapeptide containing one tyrosine as the target molecule. The reaction was readily followed by capillary electrophoresis performed at pH 8 and, using an absorption wavelength of 436 nm, the signature spectral characteristics of the nitrotyrosinylated peptide were verified on‐line. The peak appearance time for the nitrotyrosinylated peptide was more than 1 min longer than that of the starting material and a single main product was observed in contrast to the case when peroxynitrite was used as the nitrotyrosinylating reagent. Capillary electrophoresis appears to be a convenient method for the optimization of nitrotyrosinylation, examination of reaction inhibitors, and for studies of the consequences of nitrotyrosinylation, e.g. , for antibody binding and for the function of the target protein or peptide.