title: Processing, secretion, and immunoreactivity of carboxy terminally truncated dengue-2 virus envelope proteins expressed in insect cells by recombinant baculoviruses.
abstract: Two recombinant baculoviruses were constructed by inserting via the transfer vector pAcYM1 the genes coding for the structural proteins of dengue (DEN)-2 virus downstream from the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus. The two recombinants differed in truncation of 26 and 71 amino acids, respectively, in the carboxy-terminal sequence of DEN-specific envelope (E) glycoprotein. Recombinant DEN-2 E glycoproteins were processed and transported to the surface of Spodoptera frugiperda Sf9 cells infected with both viruses. We show that about one-third of the E glycoprotein minus its whole C-terminal hydrophobic anchor domain was secreted into an endoglycosidase H-resistant form. The type-specific neutralizing epitopes were conserved in the recombinant proteins as shown with a panel of monoclonal antibodies.
authors.affiliation: Laboratoire des Arbovirus, Institut Pasteur, Paris, France.
authors.forename: V
authors.initials: V
authors.lastname: Deubel
authors.forename: M
authors.initials: M
authors.lastname: Bordier
authors.forename: F
authors.initials: F
authors.lastname: Megret
authors.forename: M K
authors.initials: MK
authors.lastname: Gentry
authors.forename: J J
authors.initials: JJ
authors.lastname: Schlesinger
authors.forename: M
authors.initials: M
authors.lastname: Girard
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D001693:Biological Transport, Active; D002872:Chromosome Deletion; D003001:Cloning, Molecular / Q000379:methods; D004279:DNA, Viral / Q000235:genetics; D003716:Dengue Virus / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology; D010957:Plasmids / Q000235:genetics; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D004279:DNA, Viral; C051821:E-glycoprotein, Dengue virus type 2; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins