Peeping into human renal calcium oxalate stone matrix: characterization of novel proteins involved in the intricate mechanism of urolithiasis.
Peeping into human renal calcium oxalate stone matrix: characterization of novel proteins involved in the intricate mechanism of urolithiasis.
复制标题
DOI：
10.1371/journal.pone.0069916
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Tandon C
中科院分区：
文献类型：
--
作者：
Aggarwal KP;Tandon S;Naik PK;Singh SK;Tandon C
越来越多的患者患有尿石症，这是当今世界肾病学家面临的主要挑战之一。为了提高本病的治疗效果，肾结石形成的致病基础是需要时间的。蛋白质是人类肾结石基质的主要成分，被认为在晶体-膜相互作用、晶体生长和结石形成中具有潜在的作用，但它们在尿石症中的作用仍然不清楚。从含肾结石的人CaOx基质中分离出蛋白质。分子量为MW bbb30 kDa的蛋白质经阴离子交换层析，然后用分子筛层析。研究了纯化蛋白对CaOx成核和生长的影响，以及对草酸损伤的Madin-Darby犬肾（MDCK）肾上皮细胞的活性。通过基质辅助激光解吸/电离飞行时间（MALDI-TOF MS）鉴定蛋白质，并在MASCOT服务器上进行数据库检索。还研究了与CaOx晶体的硅分子相互作用。采用MALDI-TOF质谱法从草酸钙肾结石基质中鉴定出5种蛋白，并在MASCOT服务器上进行数据库检索，具有控制结石形成过程的能力。其中2个蛋白为启动子，2个为抑制剂，1个蛋白对CaOx的成核和生长具有抑制和促进的双重活性。进一步的分子模拟计算揭示了这些蛋白质与CaOx在分子水平上的相互作用模式。我们从人草酸钙结石基质中鉴定并鉴定了乙醇胺-磷酸胞基转移酶、Ras gtpase激活样蛋白、udp -葡萄糖糖蛋白葡萄糖基转移酶2、rms结合蛋白3A、巨噬细胞capping蛋白等在肾结石形成中起关键作用的新蛋白。因此，这些具有调节草酸钙结晶潜力的蛋白质将有助于理解和控制人类尿石症。
The increasing number of patients suffering from urolithiasis represents one of the major challenges which nephrologists face worldwide today. For enhancing therapeutic outcomes of this disease, the pathogenic basis for the formation of renal stones is the need of hour. Proteins are found as major component in human renal stone matrix and are considered to have a potential role in crystal–membrane interaction, crystal growth and stone formation but their role in urolithiasis still remains obscure. Proteins were isolated from the matrix of human CaOx containing kidney stones. Proteins having MW>3 kDa were subjected to anion exchange chromatography followed by molecular-sieve chromatography. The effect of these purified proteins was tested against CaOx nucleation and growth and on oxalate injured Madin–Darby Canine Kidney (MDCK) renal epithelial cells for their activity. Proteins were identified by Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF MS) followed by database search with MASCOT server. In silico molecular interaction studies with CaOx crystals were also investigated. Five proteins were identified from the matrix of calcium oxalate kidney stones by MALDI-TOF MS followed by database search with MASCOT server with the competence to control the stone formation process. Out of which two proteins were promoters, two were inhibitors and one protein had a dual activity of both inhibition and promotion towards CaOx nucleation and growth. Further molecular modelling calculations revealed the mode of interaction of these proteins with CaOx at the molecular level. We identified and characterized Ethanolamine-phosphate cytidylyltransferase, Ras GTPase-activating-like protein, UDP-glucose:glycoprotein glucosyltransferase 2, RIMS-binding protein 3A, Macrophage-capping protein as novel proteins from the matrix of human calcium oxalate stone which play a critical role in kidney stone formation. Thus, these proteins having potential to modulate calcium oxalate crystallization will throw light on understanding and controlling urolithiasis in humans.
登录
查看更多内容
影响因子：
8
作者：
COLOVOS, C;YEATES, TO
通讯作者：
YEATES, TO
影响因子：
5
作者：
Aggarwal, Kanu Priya;Tandon, Simran;Tandon, Chanderdeep
通讯作者：
Tandon, Chanderdeep
影响因子：
4.8
作者：
Hajjar, KA;Mauri, L;Chait, BT
通讯作者：
Chait, BT
影响因子：
4.5
作者：
Govindaraj, A;Selvam, R
通讯作者：
Selvam, R
影响因子：
4.8
作者：
Fukata, M;Kuroda, S;Kaibuchi, K
通讯作者：
Kaibuchi, K