A novel method for isolation of membrane proteins: a baculovirus surface display system.
We have developed a novel baculovirus surface display (BVSD) system for the isolation of membrane proteins. We expressed a reporter gene that encoded hemagglutinin gene fused in frame with the signal peptide and transmembrane domain of the baculovirus gp64 protein, which is displayed on the surface of BmNPV virions. The expression of this fusion protein on the virion envelope allowed us to develop two methods for isolating membrane proteins. In the first method, we isolated proteins directly from the envelope of budding BmNPV virions. In the second method, we isolated proteins from cellular membranes that had disintegrated due to viral egress. We isolated 6756 proteins. Of these, 1883 have sequence similarities to membrane proteins and 1550 proteins are homologous to known membrane proteins. This study indicates that membrane proteins can be effectively isolated using our BVSD system. Using an analogous method, membrane proteins can be isolated from other eukaryotic organisms, including human beings, by employing a host cell-specific budding virus.
Zhang
Yaozhou
Y
The Key Laboratory of Silkworm Bioreactor and Biomedicine, Zhejiang Province; Institute of Biochemistry, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China. yaozhou@zstu.edu.cn
Lv
Zhengbing
Z
Chen
Jian
J
Chen
Qin
Q
Quan
Yanping
Y
Kong
Lingyin
L
Zhang
Haihua
H
Li
Si
S
Zheng
Qingliang
Q
Chen
Jianqing
J
Nie
Zuoming
Z
Wang
Jiang
J
Jin
Yongfeng
Y
Wu
Xiangfu
X
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D012831:Bombyx; D008562:Membrane Glycoproteins / Q000235:genetics; D008565:Membrane Proteins / Q000302:isolation & purification*; D011679:Pupa / Q000821:virology; D014759:Viral Envelope Proteins / Q000302:isolation & purification; D014760:Viral Fusion Proteins / Q000235:genetics
D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins