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Investigation of tyrosine nitration and nitrosylation of angiotensin II and bovine serum albumin with electrospray ionization mass spectrometry
Author(s) -
Lee Soo Jae,
Lee Jung Rok,
Kim Young Hwan,
Park Yoon Shin,
Park Sang Ick,
Park Hyung Soon,
Kim Kwang Pyo
Publication year - 2007
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3145
Subject(s) - chemistry , mass spectrometry , tyrosine , tetranitromethane , electrospray ionization , protein mass spectrometry , sample preparation in mass spectrometry , bovine serum albumin , nitrosylation , chromatography , tandem mass spectrometry , peptide , biochemistry , nitric oxide , organic chemistry
Protein tyrosine nitration is one of the important regulatory mechanisms in various cellular phenomena such as cell adhesion, endo/exo‐cytosis of cellular materials, and signal transduction. In the present study, electrospray ionization tandem mass spectrometry (ESI‐MS/MS) with a linear ion‐trap mass spectrometer was applied for identification of nitrated proteins and localization of the modified tyrosine residues. When angiotensin II(DRVYIHPF) was nitrated in vitro with tetranitromethane (TNM), the mass spectrum showed a shift of +45 Da which corresponded to tyrosine nitration. An additional +29 Da mass shift was also detected by ESI‐MS. This differed from nitrated peptide analysis with matrix‐associated laser desorption/ionization mass spectrometry (MALDI‐MS), which showed oxygen neutral loss from the nitrated tyrosine residues upon laser irradiation. Hence the +29 Da mass shift of the nitrated peptide observed by ESI‐MS suggested the introduction of an NO group for nitrosylation of tyrosine residues. To confirm this in vitro nitrosylation on the protein level, bovine serum albumin was in vitro nitrated with TNM and analyzed by ESI‐MS/MS. As expected, +29 as well as +45 Da mass shifts were detected, and the +29 Da mass shift was found to correspond to the modification on tyrosine residues by NO. Although the chemical mechanism by which this occurs in ESI‐MS is not clear, the +29 Da mass shift could be a new potential marker of nitrosylated peptides. Copyright © 2007 John Wiley & Sons, Ltd.