肿瘤及胚胎组织特异表达蛋白65与低密度脂蛋白受体相关蛋白关联蛋白的结合  

Characterization of the interaction between low density lipoprotein receptor-related protein-associated protein 1 and the cancer and embryo expression protein 65

作　　者：靳更林[1] 张建芝[1] 苏荣[1] 刘晓颖[1] 吴健[1] 孟麟[1] 寿成超[1] 

机构地区：[1]北京大学临床肿瘤学院,北京肿瘤医院,北京100034

出　　处：《北京大学学报（医学版）》2005年第3期297-301,共5页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(30270685)项目资助;北京市科技计划重大项目(H02022002310)资助;北京大学基因中心项目的资助~~

摘　　要：目的:寻找与肿瘤及胚胎组织特异表达蛋白65(cancerandembryoexpressionprotein65,CEP65)相互结合的蛋白分子,为研究CEP65在肿瘤发生中的作用机制提供线索。方法: 以CEP65为诱饵蛋白,通过酵母双杂交方法, 筛选人胚胎组织cDNA表达文库。将获得的阳性克隆cDNA片段分别克隆到原核和哺乳细胞表达质粒,通过GSTpull down和免疫共沉淀对相互作用蛋白作进一步验证。结果: 筛选到能与CEP65相互结合的低密度脂蛋白受体相关蛋白产关联蛋白(lowdensitylipoproteinreceptor relatedprotein associatedprotein1,RAP)。将分别在原核和哺乳细胞中表达的RAP与CEP65进行GSTpull down实验,结果显示,原核表达的GST RAP可以与His CEP65相互结合,而在哺乳细胞共表达的GST CEP65和Myc RAP也能相互结合。结论:RAP可以和CEP65相结合,CEP65有可能通过与RAP相互作用,在肿瘤的浸润、转移及增生中发挥其功能。Objective:To understand the mechanism of cancer and embryo expression protein 65 (CEP65) in cancer development Methods:CEP65 was usel as a bait protein to isolate the partners of CEP65 from a human placenta cDNA library with yeast two hybrid system. The DNA fragments from positive clone were expressed prokaryotic and mammalian cells respectively, the GST pull-down experiment was performed to ascertain the binding activity. Results:After screening a human placenta cDNA library, the low density lipoprotein receptor-related protein-associated protein 1 (RAP) was isolated and shown to interact with CEP65. GST pull-down assay results indicated that His-CEP65 and GST-RAP expressed by prokaryotic system were immunoprecipitated,and so were Myc-RAP and GST-P65 expressed by mammalian cells . Conclusion: RAP can interact with CEP65 and RAP may be a candidate to mediate the function of CEP65.

关 键 词：低密度脂蛋白受体相关蛋白 特异表达蛋白 胚胎组织 关联 protein CDNA表达文库 receptor 酵母双杂交方法 相互结合 哺乳细胞 CDNA片段 相互作用蛋白 免疫共沉淀 RAP GST 蛋白分子 作用机制 肿瘤发生 诱饵蛋白 表达质粒 

分 类 号：R459.5[医药卫生—治疗学] R764.43[医药卫生—临床医学]