日本脑炎病毒受体74×10^3分子的分离与初步鉴定  

Isolation and preliminary identification of 74 ×10^3 molecule as putative Japanese encephalitis virus receptor

作　　者：任君萍[1] 雷迎峰[1] 张伟[1] 杨敬[1] 马文煜[1] 

机构地区：[1]第四军医大学微生物学教研室,西安 710032

出　　处：《中华微生物学和免疫学杂志》2009年第4期307-311,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30600526）

摘　　要：目的分离和初步鉴定日本脑炎病毒（JEV）易感细胞C6／36和Vero细胞上受体分子。方法应用免疫共沉淀（Co-IP）技术，从C6／36和Vero细胞分离与JEV结合的受体分子，做质谱分析鉴定和Western blot检测。用激光共聚焦技术（LSCM）观察候选受体分子在细胞膜上的定位及与JEV结合的情况。结果经过Co-IP反应，从C6／36、Vero细胞上分离出多个与JEV结合的分子条带。质谱分析首次鉴定出一个蛋白，即C6／36细胞上与JEV结合的相对分子质量（Mr）为74×10^3分子，为HSC70蛋白。进一步用抗HSC70抗体做Western blot，检测到从C6／36和Vero细胞膜上Co-IP分离出的74×10^3蛋白。LSCM观察到JEV吸附在C6／36细胞膜上时，HSC70蛋白与JEV共定位。结论C6／36细胞上肘，为74×10^3的HSC70可能是JEV的受体分子。Objective To isolate and identify the putative Japanese encephalitis virus（ JEV ） receptors from C6/36 and Vero cells. Methods Molecules binding with JEV were isolated from C6/36 and Vero cells by co-immunoprecipitation （Co-IP） approach, identified by mass spectrometry, and detected by Western blot. The location of putative JEV receptor on cells membrane and the binding with JEV were observed by laser scanning confocal microscopy （LSCM）. Results Several molecules binding with JEV were isolated from C6/36 and Vero cells by Co-IP, and only one molecule was identified as heat shock cognate 70 （HSC70） by mass spectrometry. Antibody against HSC70 was able to detect a 74 ×10^3 protein isolated by Co-IP from C6/36 and Vero cells membrane in Western blot assays. It was observed by LSCM that when JEV attached on the surface of C6/36 cells, JEV and HSC70 protein were co-localization. Conclusion 74 ×10^3 molecular identified as HSC70 protein from C6/36 cells may be JEV receptor.

关 键 词：日本脑炎病毒 病毒受体 免疫共沉淀 C6/36细胞 VERO细胞 

分 类 号：R3[医药卫生—基础医学]