摘要
采用蔗糖密度梯度超速离心法分离纯化高尔基体,双向凝胶电泳(2-DE)分离高尔基体蛋白质,用ImageMaster 2D软件分析所得图谱,基质辅助激光解吸离子化飞行时间质谱(MALDI—TOF MS)鉴定蛋白质点等一系列亚细胞器蛋白质组学方法建立胃癌细胞内高尔基体的蛋白图谱。结果显示分离出的纯度较高的高尔基体建立了分辨率和重复性均较好的双向电泳图谱,运用质谱技术鉴定出12个蛋白质,包括蛋白合成相关蛋白、膜融合蛋白、调节蛋白、凋亡相关蛋白、运输蛋白、细胞增殖分化相关蛋白。通过亚细胞器分离纯化,双向电泳的蛋白分离及MALDI-TOF MS蛋白鉴定分析,首次成功建立了胃癌细胞SGC7901中高尔基体的蛋白质组学技术路线,为胃癌细胞内高尔基体功能的深入研究奠定了基础。
The proteomics technique platform of Golgi apparatus isolated from gastric cancer cells was established successfully by using subcellular isolation with sucrose density gradient ultra-centrifugation,the separation of proteins of Golgi complex by 2-DE,the analysis of Electrophoresis patterns with ImageMaster 2D software,and the identification of proteins based on matrix-assisted laser desorption/ionization time of flight mass spectrometry.The results indicated that the well-resolved and reproducible 2-DE pattern of Golgi apparatus isolated from gastric cancer cells was obtained,and twelve proteins were identified,including proteins related to protein synthesis and folding,membrane fusion proteins,regulatory proteins,transportation proteins,proteins related to apoptosis,proteins related to cell proliferation.The first installment of the proteome technology of Golgi complex isolated from gastric cancer cells SGC7901 by 2-DE and MALDI-TOF MS will benefit for further investigation on the functions of Golgi complex.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2010年第2期27-31,共5页
China Biotechnology
基金
国家自然科学基金(30672431)
教育部博士点基金(20060631006)资助项目
