Identification of AcMNPV GP64-binding proteins through a combinational use of a self-biotinylated virus and the cross-linking method.
Baculoviruses are potential vectors of gene therapy for the ability to transfer gene high efficiently into mammalian cells. However, cell membrane proteins which interact with baculoviral glycoproteins have not been identified. In this study, we developed a self-biotinylated AcMNPV bearing biotinylated GP64 glycoproteins. This recombinant virus demonstrated the capability to infect insect cells and to transduct mammalian cells. Using this biotinylated virus, a protein >170Kda which could specifically interact with GP64 proteins was identified from virus transducted BHK-21 cells through cross-linking and streptavidin purification. Our study provides a useful approach for identifying cell membrane proteins that interact with baculovirus surface proteins or proteins involved in virus attachment.
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Xianliang
X
Ke
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Yuan
Y
Zhang
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Yan
Y
Liu
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. Electronic address: wanghz@wh.iov.cn.
Hanzhong
H
Wang
D000818:Animals; D019921:Biotinylation; D002460:Cell Line; D006224:Cricetinae; D003432:Cross-Linking Reagents / Q000737:chemistry; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D007668:Kidney / Q000166:cytology / Q000821:virology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D013388:Succinimides / Q000737:chemistry; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism
D003432:Cross-Linking Reagents; D008562:Membrane Glycoproteins; D011506:Proteins; D013388:Succinimides; D014759:Viral Envelope Proteins; C035759:3,3'-dithiobis(sulfosuccinimidyl propionate)
Baculovirus; Biotinylation; Cellular receptor; Cross-linking; GP64; Interaction