Application of EGFP-EGF fusions to explore mechanism of endocytosis of epidermal growth factor
Application of EGFP-EGF fusions to explore mechanism of endocytosis of epidermal growth factor作者机构：State Key Laboratory of Oncogenes and Related Genes Shanghai Cancer Institute Medical College of Fudan University Shanghai China School of Pharmacy Shanghai Jiaotong University Shanghai China
出 版 物：《Acta Pharmacologica Sinica》 (中国药理学报(英文版))
年 卷 期：2007年第28卷第1期
页 面：111-117页
核心收录：
学科分类：1001[医学-基础医学(可授医学、理学学位)] 10[医学]
基 金：Project supported by the National Basic Research Program of China(No 2004CB518802) the Science and Technology Commission of Shanghai Municipality(No 054119568)
主 题：receptor epidermal growth factor endocytosis purification expression
摘 要：Aim:To develop a simple method for monitoring protein localization of epidermalgrowth factor(EGF)in living ***:Enhanced green fluorescent protein(EGFP)was used as an autofluorescent tag to label EGF ***-PAGE andWestern blot analysis were used to detect the expression of the EGFP-tagged EGF(EGFP-EGF)*** cell-binding and internalization activity of EGFP-EGFwere analyzed by fluorescence-activated cell sorting(FACS)and confocal ***:EGFP-EGF protein was expressed in Escherichia coli and purified.A cell-binding assay demonstrated that the EGFP-EGF protein could bind effi-ciently to the cells expressing *** binding and internalization of EGFP-EGFcan be visualized even at a very low concentration under confocal *** FACS-based assay for internalization activity indicated the accumulation ofinternalized EGFP-EGF over ***,the results of the competitionassay indicated its EGFR binding *** such a method,it does notneed to label EGF with chemicals and avoid light in the experimental ***:The fusion protein EGFP-EGF has several characters including highsensitivity,stability and convenience for manipulation,and is a powerful tool forthe study of EGF endocytosis.
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