GRIM-19与HPV16E6的蛋白相互作用及位点的研究

Study of the Binding Sites and the Interation of GRIM-19 with HPV 16E6

目的探讨GRIM-19与HPV16E6的蛋白相互作用及其作用位点。方法①采用免疫共沉淀方法（CO-IP）检测GRIM-19与HPV16E6蛋白的体内结合作用；②采用GSTpulldown方法检测GRIM-19与HPV16E6蛋白的体外结合作用。结果①CO-IP方法结果显示在宫颈癌细胞株SiHa细胞中GRIM-19与HPV16E6在体内具有蛋白结合作用；②GSTpulldown方法检测结果显示GRIM-19的N端第1～35位氨基酸是与HPV16E6结合位点。结论GRIM-19与HPV16E6有蛋白结合作用，作用位点为第1～35位氨基酸残基。

Objective To explore the binding sites and the interation of GRIM-19 with HPV 16E6. Methods （1） The in vivo interation of GRIM-19 and HPV16 E6 was detected by using co-im- munoprecipitation （CO-IP） ; （2） The in vitro interation of GRIM-19 and HPV16 E6 were investigated by using GST pull down assay. Results （1） The in vivo interation of GRIM-19 and HPV16 E6 was shown by CO-IP assay in cervical cancer cell lines SiHa cells. （2） The N-terminal 1-35 amino acids of GRIM-19 were the binding sites for HPV16 E6 protein by GST pull down assay. Conclusion GRIM-19 can interact with HPV16 E6 protein, and the binding sites of GRIM-19 were the first 1- 35 amino acid residues.