title: A novel method for isolation of membrane proteins: a baculovirus surface display system.
abstract: 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.
authors.affiliation: 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
authors.forename: Yaozhou
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zhengbing
authors.initials: Z
authors.lastname: Lv
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Qin
authors.initials: Q
authors.lastname: Chen
authors.forename: Yanping
authors.initials: Y
authors.lastname: Quan
authors.forename: Lingyin
authors.initials: L
authors.lastname: Kong
authors.forename: Haihua
authors.initials: H
authors.lastname: Zhang
authors.forename: Si
authors.initials: S
authors.lastname: Li
authors.forename: Qingliang
authors.initials: Q
authors.lastname: Zheng
authors.forename: Jianqing
authors.initials: J
authors.lastname: Chen
authors.forename: Zuoming
authors.initials: Z
authors.lastname: Nie
authors.forename: Jiang
authors.initials: J
authors.lastname: Wang
authors.forename: Yongfeng
authors.initials: Y
authors.lastname: Jin
authors.forename: Xiangfu
authors.initials: X
authors.lastname: Wu
mesh_terms: 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
chemical_list: D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins