Identification of Nitrotyrosine Containing Peptides using Combined Fractional Diagonal Chromatography (COFRADIC) and Off-Line Nano-LC-MALDI |
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Authors: | Trine R Larsen Nicolai Bache Jan Bert Gramsbergen Peter Roepstorff |
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Institution: | (1) Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK 5230 Odense M, Denmark;(2) Institute of Medical Biology, University of Southern Denmark, Odense M, Denmark; |
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Abstract: | Protein nitration take place on tyrosine residues under oxidative stress conditions and may influence a number of processes
including enzyme activity, protein-protein interactions and phospho-tyrosine signalling pathways. Nitrated proteins have been
identified in a number of diseases, however, the study of these proteins has been compromised by the lack of good methods
for identifying nitrated proteins, their nitration sites and the level of nitration. Here, we present a method for identification
of nitrated peptides that allows the site specific assignment of nitration, is easy to use and reproducible, and opens up
for the possibility to quantify the level of nitration of specific peptides as function of different oxidative conditions,
namely combined fractional diagonal chromatography (COFRADIC) in combination with off-line nano-LC-MALDI. We identify six
nitrated peptides from in vitro nitrated bovine serum albumin and propose that automated COFRADIC using nano-LC and off-line MALDI-MS might be a possibility
for identification of tyrosine nitrated proteins and the nitration sites in complex samples. |
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Keywords: | |
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