Novel biomarker pipeline to probe the oxidation sites and oxidation degrees of hemoglobin in bovine erythrocytes exposed to oxidative stress
Novel biomarker pipeline to probe the oxidation sites and oxidation degrees of hemoglobin in bovine erythrocytes exposed to oxidative stress
复制标题
用于探测暴露于氧化应激的牛红细胞中血红蛋白的氧化位点和氧化程度的新型生物标志物管道
DOI：
10.1002/bmc.3612
复制
发表时间：
2016
影响因子：
1.8
通讯作者：
Xu Zhenzhen
中科院分区：
文献类型：
--
作者：
Zong Wansong;Wang Xiaoning;Yang Chuanxi;Du Yonggang;Sun Weijun;Xu Zhenzhen
研究蛋白质氧化的生物标志物可能有助于深入了解氧化应激的机制模式。本研究建立了一条以牛血红蛋白(Hb)氧化产物为生物标志物探讨其氧化机理的新途径。辐射产生的活性氧物种被用来模拟氧化应激条件来氧化牛红细胞中的Hb。经Hb提取和消化后，通过与从对照样品中提取的天然多肽的离子色谱图进行比较，对胰酶片段中的氧化肽进行了归属。随后对这些多肽的串联质谱分析证明，氧化仅限于Hb中部分暴露的氨基酸残基(例如，α-Phe36，β-MET1，β-Trp14)。对这些氧化肽的定量分析表明，目标部位的氧化程度与延长氧化剂量呈正相关，氧化过程也受残基类型的控制。与传统的蛋白质羰基分析相比，所鉴定的氧化产物是Hb氧化的可行性生物标志物，表明所提出的生物标志物管道适合为蛋白质氧化提供特异和有效的信息。版权所有©2015 John Wiley&Sons，Ltd.
Research on biomarkers for protein oxidation might give insight into the mechanistic mode of oxidative stress. In the work present here, a novel pipeline was established to probe the oxidation mechanism of bovine hemoglobin (Hb) with its oxidation products serving as the biomarkers. Reactive oxygen species generated by irradiation were used to mimic oxidative stress conditions to oxidize Hb in bovine erythrocytes. After Hb extraction and digestion, oxidized peptides in the tryptic fragments were assigned by comparison with the extracted ion chromatography spectra of native peptide from the control sample. Subsequent tandem mass spectrometry analysis of these peptides proved that oxidation was limited to partially exposed amino acid residues (α‐Phe36,β‐Met1,β‐Trp14, for instance) in Hb. Quantitation analysis on these oxidized peptides showed that oxidation degrees of target sites had positive correlations with the extended oxidation dose and the oxidation processes were also controlled by residues types. Compared with the conventional protein carbonyl assay, the identified oxidized products were feasibility biomarkers for Hb oxidation, indicating that the proposed biomarker pipeline was suitable to provide specific and valid information for protein oxidation. Copyright © 2015 John Wiley & Sons, Ltd.