title: Immobilization of diverse foreign proteins in viral polyhedra and potential application for protein microarrays.
abstract: Cypoviruses are insect viruses that produce a cytoplasmic crystalline particle called the polyhedron in which progeny virions are occluded. The virion structural protein, VP3, is implicated in the occlusion of viral particles into polyhedra. In this study, we determined the amino acid sequence of VP3 required for occlusion of viral particles into polyhedra and proposed that this sequence could be used as an immobilization signal to direct the stable incorporation of foreign proteins into polyhedra. A large-scale survey revealed that the immobilization signal could, in fact, direct the incorporation of a variety of human proteins into polyhedra. Immune reactivity and protein-protein interactions were detected on the surface of polyhedra containing immobilized foreign proteins, and these particles were shown to be highly stabilized against dehydration. We showed that these particles could be arrayed onto a glass slide by standard spotting and laser manipulation methods. Thus, this approach is well suited for protein expression, purification, and the development of protein microarrays.
authors.affiliation: Protein Crystal Corporation, Honmachi, Chuo-ku, Osaka, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Ikeda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nakazawa
authors.forename: Ai
authors.initials: A
authors.lastname: Shimo-Oka
authors.forename: Kaoru
authors.initials: K
authors.lastname: Ishio
authors.forename: Seiji
authors.initials: S
authors.lastname: Miyata
authors.forename: Yoichiroh
authors.initials: Y
authors.lastname: Hosokawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Matsumura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Masuhara
authors.forename: Serge
authors.initials: S
authors.lastname: Belloncik
authors.forename: Robert
authors.initials: R
authors.lastname: Alain
authors.forename: Naoki
authors.initials: N
authors.lastname: Goshima
authors.forename: Nobuo
authors.initials: N
authors.lastname: Nomura
authors.forename: Kenichi
authors.initials: K
authors.lastname: Morigaki
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kawai
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Kuroita
authors.forename: Bunsei
authors.initials: B
authors.lastname: Kawakami
authors.forename: Yaeta
authors.initials: Y
authors.lastname: Endo
authors.forename: Hajime
authors.initials: H
authors.lastname: Mori
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D018613:Microscopy, Confocal; D016253:Microscopy, Immunoelectron; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry*; D012087:Reoviridae / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D018411:Spodoptera
chemical_list: D011506:Proteins