Mass spectroscopic characteristics of low molecular weight proteins extracted from calcium oxalate stones: Preliminary study
Mass spectroscopic characteristics of low molecular weight proteins extracted from calcium oxalate stones: Preliminary study
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DOI：
10.1002/jcla.20214
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发表时间：
2008-01-01
影响因子：
2.7
通讯作者：
Tsai, Fuu-Jen
中科院分区：
文献类型：
--
作者：
Chen, Wen-Chi;Lai, Chein-Cheng;Tsai, Fuu-Jen
据认为，基质蛋白的边界组成可能在结石形成中发挥作用，然而，很少有关于基质蛋白在泌尿系结石的蛋白质组学研究已经进行。在这项研究中，我们从草酸钙结石中提取低分子量蛋白质，并通过质谱法测量其特征模式。共有10个结石手术切除患者的尿路结石。从结石中提取蛋白质，并通过一维电泳（十二烷基硫酸钠缓冲液[SDS]-聚丙烯酰胺凝胶电泳[SDS-PAGE]）进行鉴定。胶内消化后，样品进行分析的表面增强激光解吸电离飞行时间（SELDI-TOF）技术。从质谱数据分析肽序列。从MASCOT服务器（Matrix Science Ltd.; http://www.matrixscience.com）。从每个结石样品的SDS-PAGE中鉴定出总共三条蛋白质条带（27、18和14 kDa）。MASCOT服务器上的数据库检索（SwissProt）显示，条带1（27 kDa）中最常见的蛋白质是白细胞弹性蛋白酶前体、组织蛋白酶G前体、杀天青素前体和成髓细胞蛋白前体（EC 3.4.21.76）。条带2（18 kDa）包括钙颗粒蛋白B、嗜酸性粒细胞阳离子蛋白前体和溶菌酶C前体;条带3（14 kDa）显示嗜中性粒细胞防御素3前体、钙颗粒蛋白A、钙颗粒蛋白C和组蛋白H4。从这些蛋白质的质量模式中发现的修饰和脱酰胺化可以为基质蛋白的研究提供信息。从草酸钙结石中可以提取出各种低分子量蛋白质。这些蛋白质的特征模式及其功能有待进一步研究，以探讨它们在结石形成中的作用。
It is believed that boundary compositions of matrix proteins might play a role in stone formation; however, few proteomic studies concerning matrix proteins in urinary stones have been conducted. In this study, we extracted low molecular weight proteins from calcium oxalate stones and measured their characteristic patterns by mass spectroscopy. A total of 10 stones were surgically removed from patients with urolithiasis. Proteins were extracted from the stones and identified by one-dimensional electrophoresis (sodium dodecyl sulfate buffer [SDS]-polyacrylamide gel electrophoresis [SDS-PAGE]). After in-gel digest, samples were analyzed by the surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technique. The peptide sequences were analyzed from the data of mass spectroscopy. Proteins were identified from Database Search (SwissProt Protein Database; Swiss Institute of Bioinformatics; http://www.expasy.org/sprot) on a MASCOT server (Matrix Science Ltd.; http://www.matrixscience.com). A total of three bands of proteins (27, 18, and 14 kDa) were identified from SDS-PAGE in each stone sample. A database search (SwissProt) on a MASCOT server revealed that the most frequently seen proteins from band 1 (27 kDa) were leukocyte elastase precursor, cathepsin G precursor, azurocidin precursor, and myeloblastin precursor (EC 3.4.21.76) (leukocyte proteinase 3); band 2 (18 kDa) comprised calgranulin B, eosinophil cationic protein precursor, and lysozyme C precursor; band 3 (14kDa) showed neutrophil defensin 3 precursor, calgranulin A, calgranulin C, and histone H4. The modifications and deamidations found from the mass pattern of these proteins may provide information for the study of matrix proteins. Various lower molecular weight proteins can be extracted from calcium oxalate stones. The characteristic patterns and their functions of those proteins should be further tested to investigate their roles in stone formation.