Chinese Journal of Microbiology and Immunology 2009;29(4):307-311-

doi:10.3760/cma.j.issn.0254-5101.2009.04.005

Isolation and preliminary identification of 74×103 molecule as putative Japanese encephalitis virus receptor

Junping REN ; Yingfeng LEI ; Wei ZHANG ; Jing YANG ; Wenyu MA

Keywords

Japanese encephalitis virus; Virus receptor; Co-immunoprecipitation; C6/36 cells; Vero cells

Country

China

Language

Chinese

Abstract

Objective To isolate and identify the putative Japanese encephalitis virus (JEV) re-ceptors 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 ob-served by laser scanning confocal microscopy (LCM). 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 HSC'70 was able to detect a 74 ×103 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 HSCT0 protein were co-localization. Conclusion 74 x 103 molecular identified as HSC70 protein from C6/36 cells may be JEV receptor.