Carbonylated Plasma Proteins As Potential Biomarkers of Obesity Induced Type 2 Diabetes Mellitus
Carbonylated Plasma Proteins As Potential Biomarkers of Obesity Induced Type 2 Diabetes Mellitus
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DOI：
10.1021/pr500324y
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发表时间：
2014-11-01
影响因子：
4.4
通讯作者：
Hoffmann, Ralf
中科院分区：
文献类型：
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作者：
Bollineni, Ravi Chand;Fedorova, Maria;Hoffmann, Ralf
蛋白质羰基化是一种常见的非酶氧化翻译后修饰，通常被认为是氧化应激的生物标志物。最近的证据表明，蛋白质羰基化也与肥胖和2型糖尿病（T2 DM）有关，尽管尚未确定人血浆中羰基化的蛋白质靶点。在这项研究中，我们分析了血浆样本中的羰基化蛋白质，这些血浆样本来自瘦个体和肥胖患者（有或无T2 DM）。血浆样品用胰蛋白酶消化，羰基用O-（生物素基咔唑基甲基）羟胺衍生，通过亲和素亲和色谱富集，并通过RPC-MS/MS分析。在第二次LC-MS/MS分析中靶向潜在修饰肽的信号，以检索肽序列和修饰的残基。共鉴定出158种独特的羰基化蛋白，其中52种在所有三组的血浆样品中检测到。有趣的是，36个羰基化蛋白质仅在肥胖的T2 DM患者中检测到，而18个在两个非糖尿病组中检测到。羰基化蛋白主要来源于肝脏、血浆、血小板和内皮细胞。在功能上，它们主要参与细胞粘附、信号传导、血管生成和细胞骨架重塑。在所鉴定的羰基化蛋白质中，有几种候选蛋白质，如VEGFR-2、MMP-1、argin、MKK 4和补体C5，它们之前已经与糖尿病、肥胖症和代谢疾病相关。
Protein carbonylation is a common nonenzymatic oxidative post-translational modification, which is often considered as biomarker of oxidative stress. Recent evidence links protein carbonylation also to obesity and type 2 diabetes mellitus (T2DM), though the protein targets of carbonylation in human plasma have not been identified. In this study, we profiled carbonylated proteins in plasma samples obtained from lean individuals and obese patients with or without T2DM. The plasma samples were digested with trypsin, carbonyl groups were derivatized with O-(biotinylcarbazoylmethyl)hydroxylamine, enriched by avidin affinity chromatography, and analyzed by RPC-MS/MS. Signals of potentially modified peptides were targeted in a second LC-MS/MS analysis to retrieve the peptide sequence and the modified residues. A total of 158 unique carbonylated proteins were identified, of which 52 were detected in plasma samples of all three groups. Interestingly, 36 carbonylated proteins were detected only in obese patients with T2DM, whereas 18 were detected in both nondiabetic groups. The carbonylated proteins originated mostly from liver, plasma, platelet, and endothelium. Functionally, they were mainly involved in cell adhesion, signaling, angiogenesis, and cytoskeletal remodeling. Among the identified carbonylated proteins were several candidates, such as VEGFR-2, MMP-1, argin, MKK4, and compliment C5, already connected before to diabetes, obesity and metabolic diseases.