Processing, secretion, and immunoreactivity of carboxy terminally truncated dengue-2 virus envelope proteins expressed in insect cells by recombinant baculoviruses.
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.
Deubel
V
V
Laboratoire des Arbovirus, Institut Pasteur, Paris, France.
Bordier
M
M
Megret
F
F
Gentry
M K
MK
Schlesinger
J J
JJ
Girard
M
M
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
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D004279:DNA, Viral; C051821:E-glycoprotein, Dengue virus type 2; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins