苜蓿银纹夜蛾核型多角体病毒囊膜蛋白GP64与寄主细胞互作蛋白的初步鉴定

Identification of Host Cell Protein Interacting with Envelope Protein GP64 of Autographa californica Multicapsid Nucleopolyhedrovirus

鉴定杆状病毒与宿主昆虫细胞间的互作蛋白,对于进一步研究病毒受体的特性与功能,理解病毒与宿主细胞的相互作用关系具有重要意义。为了阐明苜蓿银纹夜蛾核型多角体病毒(Ac MNPV)出芽型病毒粒子(BV)囊膜蛋白GP64是否与宿主细胞表面蛋白存在互作,利用蛋白酶K消化处理Tn细胞系的细胞膜蛋白,结果表明BV不能感染经300μg/m L蛋白酶K消化处理的Tn细胞,免疫荧光检测BV病毒粒子不能吸附于Tn细胞膜上。利用病毒覆盖蛋白印迹实验(VOPBA)及质谱初步鉴定的结果显示,Tn细胞膜上的翻译控制肿瘤蛋白(TCTP)是与BV囊膜蛋白GP64互作的候选蛋白。

Identification of insect cell proteins interacting with baculovirus is of significant importance for further explora- tion on characteristics and functions of virus receptors and for understanding the interaction between virus and host cell. To identify cell membrane proteins interacting with envelope protein GP64 of budded virus （BV） of Autographa californica multicapsid nucleopolyhedrovirus （AcMNPV）, cell membrane proteins of Tn cells were digested with protease K. The re- sult revealed that BV could not infect Tn cells that had been digested with 300 μg/mL protease K. Immunofluorescence assay displayed that BV virus particles could not attach onto Tn cell membrane. Using virus overlay protein binding assay （VOPBA） and mass spectrometry, translationally-controlled tumor protein （TCTP） of Tn cell membrane was preliminarily identified as the candidate protein interacting with envelope protein GP64 of BV.