Tandem mass spectrometry for the study of glyoxal-derived advanced glycation end-products (AGEs) in peptides.
Tandem mass spectrometry for the study of glyoxal-derived advanced glycation end-products (AGEs) in peptides.
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用于研究肽中乙二醛衍生的晚期糖基化终产物 (AGE) 的串联质谱法。
DOI：
10.1002/rcm.6753
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发表时间：
2014
期刊：
影响因子：
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通讯作者：
O'Connor,PeterB
中科院分区：
文献类型：
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作者：
Lopez-Clavijo,AndreaF;Duque-Daza,CarlosA;O'Connor,PeterB
基本原理称为糖化的翻译后修饰会影响肽和蛋白质的生理特性。糖化在高血糖期间尤其重要，其中会产生 α-二羰基化合物。这些化合物与蛋白质反应生成α-二羰基衍生的糖基化产物，这些产物与糖尿病并发症如肾病、视网膜病和神经病等相关。这些α-二羰基化合物之一是乙二醛，也称为乙二醛。因此，通过电子捕获解离 (ECD) 和碰撞激活解离 (CAD) 研究乙二醛与蛋白质/肽的结合。 方法在拟生理和 MeOH/H2O (50:50) 条件下，使含有 1 个赖氨酸和 1 个精氨酸的易受糖化影响的乙酰化和非乙酰化十一肽与乙二醛反应。使用 12 Tesla 傅里叶变换离子回旋共振质谱 (FTICRMS) 通过 ECD 和 CAD 裂解两种类型的乙二醛衍生 AGE。 结果在不同反应条件下与乙二醛的反应显示添加了 C2O 和 C2H2O2，这对应于肽质量分别净增加了 39.9949 Da 和 58.0055 Da。使用 ECD 和 CAD 分配结合位点的误差小于 1 ppm。结果表明，两种类型的乙二醛衍生 AGE 均在位于 3 位的精氨酸侧链上形成。 结论 使用 FTICRMS 对含有一个精氨酸-一个赖氨酸的肽中乙二醛衍生 AGE 的类型和结合位点进行了研究。肽质量的两个净质量添加被指定为 C2O 和 C2H2O2，它们位于精氨酸侧链。此外，在肽中观察到的这些质量添加（C2O 和 C2H2O2）不受不同反应条件的影响。版权所有 © 2013 约翰·威利父子有限公司
RATIONALEThe post‐translational modification known as glycation affects the physiological properties of peptides and proteins. Glycation is particularly important during hyperglycaemia where α‐dicarbonyl compounds are generated. These compounds react with proteins to generate α‐dicarbonyl‐derived glycation products, which are correlated with diabetic complications such as nephropathy, retinopathy, and neuropathy, among others. One of these α‐dicarbonyl compounds is ethanedial, also known as glyoxal. Thereby, glyoxal binding to protein/peptides is studied by electron capture dissociation (ECD) and collisionally activated dissociation (CAD).METHODSAcetylated and non‐acetylated undecapeptides containing one lysine and one arginine susceptible of glycation were reacted with glyoxal under pseudo‐physiological and MeOH/H2O (50:50) conditions. Two types of glyoxal‐derived AGEs were fragmented by ECD and CAD using 12 Tesla Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS).RESULTSReaction with glyoxal under different reaction conditions showed the addition of C2O and C2H2O2, which corresponded to a net increase on the peptide mass of 39.9949 Da and 58.0055 Da, respectively. The binding site was assigned within an error <1 ppm, using ECD and CAD. The results indicated that both types of glyoxal‐derived AGEs are formed at the side chain of arginine located in position 3.CONCLUSIONSTypes and binding sites of glyoxal‐derived AGEs were investigated in peptides containing one arginine‐one lysine using FTICRMS. Two net mass additions to the mass of the peptide were assigned as C2O and C2H2O2, which were located at the arginine side chain. In addition, these mass additions (C2O and C2H2O2) observed in the peptides were unaffected by different reaction conditions. Copyright © 2013 John Wiley & Sons, Ltd.