Proteomic analysis of proteins selectively associated with hydroxyapatite, brushite, and uric acid crystals precipitated from human urine.
Proteomic analysis of proteins selectively associated with hydroxyapatite, brushite, and uric acid crystals precipitated from human urine.
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DOI：
10.1021/pr1006312
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发表时间：
2010-08
影响因子：
4.4
通讯作者：
L. Thurgood;R. Ryall
中科院分区：
文献类型：
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作者：
L. Thurgood;R. Ryall
本研究的目的是比较从同一尿样中沉淀的羟基磷灰石(HA)、刷石(BR)和尿酸(UA)晶体内的蛋白质谱。HA、BR和UA结晶在两个不同的场合从相同的健康尿液中沉淀出来。洗涤晶体以去除表面结合的蛋白质，并用傅里叶变换红外光谱(FTIR)、场发射扫描电子显微镜(FESEM)和能量色散X射线分析(EDAX)确定了其组成。用SDS-PAGE对脱矿晶体提取物的蛋白质含量进行直观比较，并用LC-MS/MS对其进行分析。HA由纳米颗粒和有机物质组成，有机物质在BR和UA晶体中不存在。3种矿物的单个蛋白质的数量和类型不同：在HA晶体提取物中检测到45种蛋白质，在BR晶体中检测到77种蛋白质，包括一些被视为方法学污染物的角蛋白。剔除角蛋白后，HA和BR晶体共有21种蛋白质。在尿酸提取物中鉴定出7种非角蛋白。在HA和BR晶体提取物中持续检测到的几种蛋白质以前被认为与肾结石疾病有关，包括骨桥蛋白、凝血酶原、蛋白S100A9(钙粒蛋白B)、α间抑制物、α1-微球蛋白bikunin(AMBP)、硫酸乙酰肝素蛋白多糖和Tamm-Horsfall糖蛋白，所有这些都是强钙结合蛋白。我们的结论是，蛋白质与健康尿液中形成的HA、BR和UA晶体的结合是有选择性的，健康尿液中存在的众多蛋白质中只有几个可能在预防结石发病方面发挥任何重要作用。
The aim of this study was to compare the intracrystalline protein profiles of hydroxyapatite (HA), brushite (BR), and uric acid (UA) crystals precipitated from the same urine samples. HA, BR, and UA crystals were precipitated on two different occasions from the same pooled healthy urine. Crystals were washed to remove surface-bound proteins, and their composition was confirmed using Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM) coupled with energy dispersive X-ray analysis (EDAX). SDS-PAGE was used for visual comparison of the protein content of the demineralised crystal extracts, which were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). HA comprised nanosized particles interspersed with organic material, which was absent from the BR and UA crystals. The number and type of individual proteins differed between the 3 minerals: 45 proteins were detected in the HA crystal extracts and 77 in the BR crystals, including a number of keratins, which were regarded as methodological contaminants. After excluding the keratins, 21 proteins were common to both HA and BR crystals. Seven nonkeratin proteins were identified in the UA extracts. Several proteins consistently detected in the HA and BR crystal extracts have been previously implicated in kidney stone disease, including osteopontin, prothrombin, protein S100A9 (calgranulin B), inter-α-inhibitor, α1-microglobulin bikunin (AMBP), heparan sulfate proteoglycan, and Tamm-Horsfall glycoprotein, all of which are strong calcium binders. We concluded that the association of proteins with HA, BR, and UA crystals formed in healthy urine is selective and that only a few of the numerous proteins present in healthy urine are likely to play any significant role in preventing stone pathogenesis.