利用双向电泳技术分析正常大鼠肾小球蛋白质组方法的建立  

Establishment of Method on Analyzing the Proteomics of Rat Glomeruli by Two-dimensional Gel Electrophoresis

作　　者：吕杨[1] 谢院生[1] 张术佳[1] 李晓帆[1] 谢振声[2] 杨福全[2] 陈香美[1] 

机构地区：[1]解放军总医院肾病科,全军肾脏病研究所暨重点实验室,北京100853 [2]中国科学院生物物理研究所,北京100101

出　　处：《中国中西医结合肾病杂志》2010年第3期197-199,共3页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金重点资助项目(No.30630033)

摘　　要：目的:利用双向电泳技术分析大鼠肾小球蛋白质,以建立肾小球蛋白质组分析的方法。方法:选取体重约200g的雄性Wistar大鼠,分离肾小球并提取蛋白质。使用24cm、pH3-11的IPG胶条对蛋白质进行一相等电聚焦,采用12.5%聚丙烯酰胺凝胶进行二相分离。Decyder软件分析凝胶上的蛋白质点,切取1个蛋白质点,经过胰蛋白酶消化后,使用飞行质谱获取肽指纹谱,并用Mascot软件分析。结果:纯化的肾小球纯度在90%以上,在双向电泳的凝胶上,共发现1240个肾小球蛋白质;切取的蛋白质点经鉴定为CAPZA2蛋白质,Mascot得分为137分(阈值为61分)。结论:成功建立了双向电泳分析大鼠肾小球蛋白质组的方法。Objective：To establish the method of analyzing the proteomics of rat glomeruli by Two-dimensional gel electrophoresis （2-DE）.Methods：The glomeruli protein isolated from 200 g wistar rat was separated by 24 cm pH 3-11 IPG strip and 12.5% polyacrylamide gel.The number of protein spot was calculated by Decyder software.The protein spot was digested by tripsin and the peptide mapping fingerprint was obtained by Maldi-tof.The results were analyzed by Mascot software.Results：The purity of glomeruli was more than 90%.There were 1 240 spots being found in 2-DE map.The protein spot was identified as capping protein muscle Z-line alpha 2 （CAPZA2） protein with the value of 137 calculated by mascot （threshold was 61）.Conclusion：The method of analyzing the rat glomeruli by 2-DE map is established successfully.

关 键 词：双向电泳 肾小球 CAPZA2 

分 类 号：R692[医药卫生—泌尿科学]