Mass spectrometric identification of in vivo nitrotyrosine sites in the human pituitary tumor proteome.
Mass spectrometric identification of in vivo nitrotyrosine sites in the human pituitary tumor proteome.
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DOI：
10.1007/978-1-59745-562-6_10
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发表时间：
2009
影响因子：
--
通讯作者：
X. Zhan;D. Desiderio
中科院分区：
文献类型：
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作者：
X. Zhan;D. Desiderio
The chemically stable tyrosine nitration of a protein involves the addition of a nitro group (–NO2) to the phenolic ring of a tyrosine residue, which may be associated with nervous system physiological and pathological processes. Identification of nitrotyrosine sites on a protein could clarify the functional significance of the modification. Due to the rarity of nitrotyrosine sites in a proteome, tandem mass spectrometry, coupled with different techniques that isolate and enrich nitrotyrosine-containing proteins from a pituitary proteome, is currently the most effective method for site identification. Commercially available nitrotyrosine polyclonal/monoclonal antibodies enable one to detect nitrotyrosine-containing proteins in a two-dimensional gel electrophoresis (2DGE) map, and to preferentially enrich nitrotyrosine-containing proteins with immunoprecipitation. Our present protocols have integrated different isolation/enrichment techniques (2DGE; Western blots; nitrotyrosine immunoaffinity precipitation) and two different tandem mass spectrometry methods (MALDI-MS/MS; ESI-MS/MS) to determine the amino acid sequence of nitrotyrosine-containing peptides that derive from nitrated proteins. Bioinformatics tools are then used to correlate nitrotyrosine sites with a functional domain/motif in order to understand the relationship between tyrosine nitration and the structural/functions of proteins.
