Investigation of tyrosine nitration and nitrosylation of angiotensin II and bovine serum albumin with electrospray ionization mass spectrometry
Article first published online: 27 JUL 2007
Copyright © 2007 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 21, Issue 17, pages 2797–2804, 15 September 2007
How to Cite
Lee, S. J., Lee, J. R., Kim, Y. H., Park, Y. S., Park, S. I., Park, H. S. and Kim, K. P. (2007), Investigation of tyrosine nitration and nitrosylation of angiotensin II and bovine serum albumin with electrospray ionization mass spectrometry. Rapid Commun. Mass Spectrom., 21: 2797–2804. doi: 10.1002/rcm.3145
- Issue published online: 27 JUL 2007
- Article first published online: 27 JUL 2007
- Manuscript Accepted: 17 JUN 2007
- Manuscript Revised: 14 JUN 2007
- Manuscript Received: 22 FEB 2007
- Korean Health 21 R&D Project, Ministry of Health & Welfare, Republic of Korea. Grant Number: A060769
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 invitro 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.