Proteomic Analysis after Sequential Extraction of Matrix Proteins in Urinary Stones Composed of Calcium Oxalate Monohydrate and Calcium Oxalate Dihydrate
Proteomic Analysis after Sequential Extraction of Matrix Proteins in Urinary Stones Composed of Calcium Oxalate Monohydrate and Calcium Oxalate Dihydrate
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DOI：
10.2116/analsci.31.935
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发表时间：
2015-09-01
影响因子：
1.6
通讯作者：
Horie, Shigeo
中科院分区：
文献类型：
--
作者：
Kaneko, Kiyoko;Nishii, Shin-ichiro;Horie, Shigeo
在这项研究中，我们进行了蛋白质组学分析后，顺序蛋白质提取的一水草酸钙（COM）和二水草酸钙（COD）的尿结石，以确定特定的基质蛋白，根据晶体成分的石头。对13例尿结石进行X射线和红外光谱分析后，依次用KCl、甲酸、盐酸胍和EDTA提取基质蛋白，然后进行SDS-电泳，然后进行液相色谱-质谱/质谱（LC-MS/MS）。提取的蛋白质的电泳图谱不同于COM和COD结石。LC-MS/MS鉴定了65种蛋白质，其中许多是细胞血浆蛋白，并且无论晶体组分如何都经常被检测到。然而，在COM结石的最终EDTA组分中检测到6种蛋白质（蛋白Z、蛋白S、凝血酶原、骨桥蛋白、脂肪酸结合蛋白5和泛素）。这些蛋白质参与凝血过程或骨代谢，因此它们所起的作用特别令人感兴趣。
In this study, we performed proteomic analysis following sequential protein extraction on calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) urinary stones to determine the specific matrix proteins according to the crystal components of the stones. After X-ray and IR analysis of 13 urinary stones, matrix proteins were sequentially extracted with KCl, formic acid, guanidine-HCl, and EDTA, before SDS-electrophoresis followed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The electrophoretic patterns of the extracted proteins differed from that of COM and COD stones. LC-MS/MS identified 65 proteins, of which many were cellular plasma proteins, and were frequently detected regardless of the crystal components. However, 6 proteins (protein Z, protein S, prothrombin, osteopontin, fatty acid binding protein 5, and ubiquitin) were detected in the final EDTA fractions of COM stones. These proteins are involved in the coagulation process or osteometabolism, and thus the roles they play are of particular interest.