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Site‐specific proteomic approach for study protein S‐nitrosylation
Author(s) -
Ding ShiJian,
Liu Miao
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.959.2
Subject(s) - chemistry , s nitrosylation , cysteine , nitric oxide , tandem mass spectrometry , mass spectrometry , nitrosylation , posttranslational modification , biochemistry , tandem mass tag , proteomics , biophysics , quantitative proteomics , chromatography , biology , enzyme , organic chemistry , gene
Protein S‐nitrosylation, the covalent attachment of nitric oxide (NO) to cysteine residues, is an emerging post‐translational modification (PTM) which regulates a large variety of cellular functions and signalling events. Characterization of S‐nitrosylated proteins and their modification sites is essential to understand the action of NO in biological systems. Here we present a novel and robust method for the identification of protein S‐nitrosylation sites in complex protein mixtures. The approach utilizes the Cysteinyl affinity resin to selectively enrich S‐nitrosylated peptides. The enriched samples were analyzed by nanoscale liquid chromatography tandem mass spectrometry on LTQ‐Orbitrap mass spectrometer. We applied this approach to MDA‐MB‐231 cells treated with Angeli's salt. Angeli's salt has been shown to inhibit breast cancer tumor growth and angiogenesis. A total of 177 S‐nitrosylation sites were identified and an S‐nitrosylation motif was revealed in our study. The 177 sites are significantly more than the number reported by previous methods, demonstrating the efficiency of our approach.