小鼠肝细胞核磷酸化蛋白质组的提取和双向电泳分离  

Purification and Two-Dimensional Electrophoresis Analysis of Nuclear Phosphoproteins of Mouse Liver Cells

作　　者：夏高晓[1] 李红梅[1] 陈丽[1] 赵明哲[1] 胡水旺[1] 王继刚[1] 孙明晶[1] 邓鹏[1] 刘靖华[1] 姜勇[1] 

机构地区：[1]南方医科大学病理生理学教研室,广东省蛋白质组学重点实验室,广州510515

出　　处：《热带医学杂志》2009年第5期487-489,共3页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.30572151;No.30670828);广东省自然科学基金(No.05004730);广州市科技计划项目(No.2007J1-C0301);国家自然科学基金委员会-广东省人民政府自然科学联合基金重点项目(No.U0632004);教育部长江学者和创新团队发展计划项目(No.IRT0731)

摘　　要：目的提取小鼠肝细胞核磷酸化蛋白并利用双向凝胶电泳分离小鼠肝细胞核磷酸化蛋白。方法提取小鼠肝细胞核蛋白并利用金属磷酸盐亲和层析树脂纯化出磷酸化蛋白后,进行一维等电聚焦分离和二维聚丙烯酰胺凝胶电泳分离,并挑选其中一个蛋白斑点进行质谱分析。结果成功提取了肝细胞核磷酸化蛋白,通过双向凝胶电泳分离技术成功建立了小鼠肝细胞核磷酸化蛋白质组图谱。质谱鉴定结果证实被挑选蛋白斑点是小鼠核磷酸化蛋白。结论磷酸化蛋白纯化技术结合二维凝胶电泳分离技术是研究肝细胞核磷酸化蛋白质组的有效方法,为进一步全面研究和鉴定小鼠肝细胞核磷酸化蛋白的功能打下了基础。Objective To analyze the nuclear phosphoproteins from mouse liver cells by two-dimensional electrophoresis. Methods Nuclear phosphoproteins were enriched by phosphate metal affinity chromatography resin （PMAC）. The isolated proteins were then analyzed by 2D electrophoresis. Results Nuclear phosphoproteins were successfully purified from mouse liver cells. One of the spots was identified as the nuclear phosphoprotein by mass spectroscopy. Conclusion An effective approach was developed to identify nuclear phosphoproteins. This method would provide a solid foundation for the further studies on the functions of nuclear phosphoproteins.

关 键 词：细胞核磷酸化蛋白 磷酸化蛋白质组 双向凝胶电泳 肝脏 质谱 

分 类 号：Q51[生物学—生物化学] Q503