Novel antilithiatic cationic proteins from human calcium oxalate renal stone matrix identified by MALDI-TOF-MS endowed with cytoprotective potential: An insight into the molecular mechanism of urolithiasis
Novel antilithiatic cationic proteins from human calcium oxalate renal stone matrix identified by MALDI-TOF-MS endowed with cytoprotective potential: An insight into the molecular mechanism of urolithiasis
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DOI：
10.1016/j.cca.2012.10.040
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发表时间：
2013-01-16
影响因子：
5
通讯作者：
Tandon, Chanderdeep
中科院分区：
文献类型：
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作者：
Aggarwal, Kanu Priya;Tandon, Simran;Tandon, Chanderdeep
背景：迄今为止，尚无关于具有抗石活性的阳离子蛋白的实质性研究。我们探索了人类草酸钙（CaOx）结石中存在的抗锂离子阳离子蛋白，并研究了它们与硅中草酸钙晶体的分子相互作用。方法：从人肾结石CaOx基质中分离蛋白质。用阳离子交换层析法和分子筛层析法对分子量为MW bbb3kda的蛋白质进行了分析。这些纯化的阳离子蛋白对CaOx成核和生长的影响以及对草酸损伤的MDCK细胞的活性进行了测试。用MALDI-TOF ms鉴定了蛋白质，并研究了它们与硅晶COM的分子相互作用。结果：基于MASCOT服务器数据库检索，MALDI-TOF MS鉴定出3种抗锂离子阳离子蛋白，分别为组蛋白-赖氨酸n -甲基转移酶、内向整流K通道和蛋白Wnt-2，分子量分别为53、44和42 kDa。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。结论：组蛋白-赖氨酸n -甲基转移酶、内向整流K通道和蛋白Wnt-2是新的抗石蛋白，在肾脏功能中起重要作用，并与多种肾脏疾病有关。(c) 2012 Elsevier B.V.版权所有
Background: No substantial work has been conducted to date in context to cationic proteins with antilithiatic activity. We explored the antilithiatic cationic proteins present in human calcium oxalate (CaOx) stones and also examined their molecular interactions with calcium oxalate crystals in silico.Methods: Proteins were isolated from the matrix of human CaOx containing kidney stones. Proteins having MW>3 kDa were subjected to cation exchange chromatography followed by molecular-sieve chromatography. The effect of these purified cationic proteins was tested against CaOx nucleation and growth and on oxalate injured MDCK cells for their activity. Proteins were identified by MALDI-TOF MS. Molecular interaction studies with COM crystals in silico were also investigated.Results: Three antilithiatic cationic proteins were identified as histone-lysine N-methyltransferase, inward rectifier K channel and protein Wnt-2 (MW similar to 53, similar to 44, and similar to 42 kDa respectively) by MALDI-TOF MS based on database search with MASCOT server. Further molecular modeling calculations revealed the mode of interaction of these proteins with CaOx at the molecular level.Conclusion: We identified histone-lysine N-methyltransferase, inward rectifier K channel and protein Wnt-2 as novel antilithiatic proteins which play a vital role in the kidney function and have been associated with various kidney diseases. (c) 2012 Elsevier B.V. All rights reserved.