Simultaneous measurement of protein-bound 3-chlorotyrosine and homocitrulline by LC-MS/MS after hydrolysis assisted by microwave: Application to the study of myeloperoxidase activity during hemodialysis
Simultaneous measurement of protein-bound 3-chlorotyrosine and homocitrulline by LC-MS/MS after hydrolysis assisted by microwave: Application to the study of myeloperoxidase activity during hemodialysis
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DOI：
10.1016/j.talanta.2012.06.044
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发表时间：
2012-09-15
期刊：
影响因子：
6.1
通讯作者：
Boudjeltia, Karim Zouaoui
中科院分区：
文献类型：
--
作者：
Delporte, Cedric;Franck, Thierry;Boudjeltia, Karim Zouaoui
高程度的尿毒症在终末期肾病患者中很常见，并且与慢性炎症和心血管疾病的发展有关。在不可能进行移植的情况下，尿毒症可以通过血液透析来减轻，尽管重复干预涉及肾功能丧失，部分原因是慢性炎症和/或氧化应激过程。在这种情况下，它已经表明，髓过氧化物酶（MPO）可以有助于血液透析过程中的氧化应激和心血管风险。在血液透析过程中，由于MPO活性而导致的蛋白质损伤从未被评估过，尽管其两种反应产物3-氯酪氨酸和高瓜氨酸是令人感兴趣的。事实上，第一个是MPO活性的特异性产物，第二个的形成可以由MPO催化。为了分析血浆蛋白中的这些产物，开发了一种全水解方法，然后进行液相色谱质谱分析。对不同的水解条件进行了试验，并对完全水解和人工氯化的最佳工艺进行了评估。最后，该方法用于分析3-氯酪氨酸和高瓜氨酸在血液透析期间的血浆蛋白质在15例患者和数据相关的MPO浓度和活性的测量。观察到MPO活性和蛋白结合的3-氯酪氨酸均增加，突出了MPO参与血液透析期间的氧化应激，并进一步证明了血液透析与心血管疾病之间的联系。(C)2012爱思唯尔有限公司版权所有。
A high degree of uremia is common in patients with end-stage renal disease and has been linked to the development of chronic inflammation and cardiovascular diseases. In conditions where transplantation is not possible, uremia can be reduced by hemodialysis although the repeated interventions have been implicated in loss of renal function, partially as a result of chronic inflammation and/or oxidative stress processes. In this context, it has been suggested that myeloperoxidase (MPO) can contribute to the oxidative stress during hemodialysis and to the cardiovascular risk. Protein damages due to MPO activity have never been assessed during hemodialysis although two of its reaction products, 3-chlorotyrosine and homocitrulline, are of interest. Indeed, the first one is a specific product of MPO activity and the formation of the second one could be catalyzed by MPO. In order to analyze these products in plasma proteins, a total hydrolysis method followed by liquid chromatography mass spectrometry analysis was developed. Different conditions of hydrolysis were tested and the optimized procedure was assessed for complete hydrolysis and artifactual chlorination. Finally, the method was used for analyzing 3-chlorotyrosine and homocitrulline in plasma proteins during a hemodialysis session in fifteen patients and data were related to measurements of MPO concentration and activity. Both increases in MPO activity and protein-bound 3-chlorotyrosine were observed, highlighting the involvement of MPO in oxidative stress during hemodialysis and further demonstrating the link between hemodialysis and cardiovascular diseases. (C) 2012 Elsevier B.V. All rights reserved.