Aggarwal Anshu, Tandon Simran, Singla Surinder Kumar, Tandon Chanderdeep
Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat- 173234, Solan, India.
Protein Pept Lett. 2012 Aug;19(8):812-9. doi: 10.2174/092986612801619552.
Adhesion of calcium oxalate (CaOx) crystals to kidney cells is a key event in kidney stones associated with marked hyperoxaluria. As the propensity of stone recurrence and persistent side effects are not altered by surgical techniques available, phytotherapeutic agents could be useful as an adjuvant therapy. The present study is aimed at examining the antilithiatic potency of the protein biomolecules of Tribulus terrestris, a plant which is a common constituent of herbal marketed preparations to treat urolithiasis. Various biochemical methods with mass spectrometry were used to purify and characterize the purified protein. The protective potency of the protein was tested on the oxalate induced injury on renal epithelial cell lines (NRK 52E). An antilithiatic protein having molecular weight of ~ 60kDa was purified. This purified protein showed similarities with Carotenoid cleavage dioxygenase 7 (CCD7) of Arabidopsis thaliana after matching peptide mass fingerprints in MASCOT search engine. An EF hand domain was identified in CCD7 by SCAN PROSITE. Presence of an EF hand domain, a characteristic feature of calcium binding proteins and a role in the synthesis of retinol which is transported by retinol binding protein, a protein found in kidney stone matrix; of CCD7 support the role of TTP as an antilithiatic protein. The protective potency of TTP on NRK 52E was quite comparable to the aqueous extract of cystone. Our findings suggest that this purified protein biomolecule from Tribulus terrestris could open new vista in medical management of urolithiasis.
草酸钙（CaOx）晶体与肾细胞的黏附是与显著高草酸尿症相关的肾结石形成中的关键事件。由于现有手术技术无法改变结石复发倾向和持续的副作用，植物治疗剂可能作为辅助治疗手段发挥作用。本研究旨在检验刺蒺藜的蛋白质生物分子的抗结石效力，刺蒺藜是市售治疗尿石症草药制剂的常见成分。采用各种生化方法并结合质谱来纯化和表征纯化后的蛋白质。在肾上皮细胞系（NRK 52E）上测试了该蛋白质对草酸盐诱导损伤的保护效力。纯化得到一种分子量约为60kDa的抗结石蛋白质。在MASCOT搜索引擎中匹配肽质量指纹后，该纯化蛋白质显示出与拟南芥的类胡萝卜素裂解双加氧酶7（CCD7）相似。通过SCAN PROSITE在CCD7中鉴定出一个EF手型结构域。CCD7中存在EF手型结构域，这是钙结合蛋白的一个特征，并且在视黄醇的合成中起作用，视黄醇由视黄醇结合蛋白转运，视黄醇结合蛋白是在肾结石基质中发现的一种蛋白质；这支持了刺蒺藜蛋白作为抗结石蛋白的作用。刺蒺藜蛋白对NRK 52E的保护效力与胱氨酸片的水提取物相当。我们的研究结果表明，从刺蒺藜中纯化得到的这种蛋白质生物分子可能为尿石症的医学管理开辟新的前景。
