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人肾组织磷酸化蛋白质组二维凝胶电泳的建立与优化
刘亚伟,戴兵,梅长林,沙伟,张岩,熊锡山,LIUYa-wei,DAIBing,MEIChang-lin,SHAWei,ZHANGYan,XIONGXi-shan
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摘要:
目的:利用二维凝胶电泳法分离人肾组织磷酸化蛋白质组.方法:利用金属磷酸盐亲和层析树脂富集人肾组织磷酸化蛋白.样品浓缩除盐后,进行一维等电聚焦分离和二维SDS电泳分离提取人肾组织磷酸化蛋白.结果:成功提取了人肾组织磷酸化蛋白,并得到了磷酸化蛋白的双向凝胶电泳图谱.结论:磷酸化蛋白富集技术与二维凝胶电泳技术的结合是研究磷酸化蛋白质组的有效方法,为进一步研究人肾组织磷酸化蛋白奠定基础.
关键词:  蛋白质组学、磷酸化蛋白质组、二维凝胶电泳
DOI:10.3724/SP.J.1008.2006.00603
投稿时间:2006-04-11修订日期:2006-05-31
基金项目:国家自然科学基金重点项目(30330640);国家自然科学基金面上项目(30271523);国家自然科学基金(30576867).
Establishment and optimization of two-dimensional gel electrophoresis for separation of human kidney phosphoproteome
刘亚伟,戴兵,梅长林,沙伟,张岩,熊锡山,LIU Ya-wei,DAI Bing,MEI Chang-lin,SHA Wei,ZHANG Yan,XIONG Xi-shan
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Abstract:
Objective:To separate human kidney phosphoproteome by two-dimensional gel electrophoresis. Methods: The phosphorylated proteins from human kidney tissues were enriched with phosphate metal affinity chromatography (PMAC) resin. After being concentrated and desalted, the samples were separated by isoelectric focusing on first dimension and SDS electrophoresis on second dimension. Results: The phosphorylated proteins were successfully extracted from human kidney tissues and were separated by two-dimensional gel electrophoresis. Conclusion.. Phosphoprotein enrichment technique combined with two-dimensional gel electrophoresis is an effective approach to study phosphoproteome, laying a foundation for further investigation of human kidney phosphoproteins
Key words:  proteomics  phosphoproteome  two-dimensional gel electrophoresis