Efficient identification and quantification of peptides containing nitrotyrosine by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after derivatization.

Efficient identification and quantification of peptides containing nitrotyrosine by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after derivatization.
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衍生化后通过基质辅助激光解吸/电离飞行时间质谱法有效鉴定和定量含有硝基酪氨酸的肽。

DOI:
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发表时间:
2010
影响因子:
1.7
通讯作者:
K. Kohda
K. Kohda
中科院分区:
医学4区
文献类型:
--
作者:
H. Tsumoto;R. Taguchi;K. Kohda

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酪氨酸残基处的蛋白质硝化继续形成3-硝基酪氨酸;该产物今天被用作硝化应激的生物标志物。我们已经开发出一种有效的方法,其中确定硝化肽和定量蛋白质硝化水平在不同的生物样品的含硝基酪氨酸的肽和质谱的化学衍生的组合。我们的战略包括:1)通过用(13)C(0)/(13)C(4)-或D(0)/D(6)-乙酸酐乙酰化来保护赖氨酸残基的N-末端胺和ε-胺; 2)用连二亚硫酸钠将硝基酪氨酸还原成氨基酪氨酸;和3)用1-(6-甲基[D(0)/D(3)]烟酰氧基)琥珀酰亚胺衍生化氨基酪氨酸。我们的方法的效用被证明与硝基酪氨酸含血管紧张素II和牛血清白蛋白作为模型化合物。
Protein nitration at tyrosine residues proceeds to form 3-nitrotyrosine; this product is today used as a biomarker of nitrative stress. We have developed an efficient method with which to identify nitrated peptides and to quantify protein nitration levels in different biological samples by a combination of the chemical derivatization of nitrotyrosine-containing peptides and mass spectrometry. Our strategy includes: 1) the protection of both N-terminal amines and epsilon-amines of lysine residues by acetylation with (13)C(0)/(13)C(4)- or D(0)/D(6)-acetic anhydride; 2) the reduction of nitrotyrosine to aminotyrosine with sodium hydrosulfite; and 3) the derivatization of aminotyrosine with 1-(6-methyl[D(0)/D(3)]nicotinoyloxy)succinimide. The utility of our method is demonstrated with nitrotyrosine-containing angiotensin II and bovine serum albumin as the model compounds.
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