title: Altering the surface properties of baculovirus Autographa californica NPV by insertional mutagenesis of the envelope protein gp64.
abstract: The envelope protein gp64 of the baculovirus Autographa californica nuclear polyhedrosis virus is essential for viral entry into insect cells, as the glycoprotein both mediates pH-dependent membrane fusion and binds to host cell receptors. Surface modification of baculovirus particles by genetic engineering of gp64 has been demonstrated by various strategies and thus has become an important and powerful tool in molecular biology. To improve further the presentation of peptides on the surface of baculovirus particles, several insertion sites within the gp64 envelope protein were selected by their theoretical maximum surface probability and investigated for efficient peptide presentation. The ELDKWA peptide of the gp41 of HIV-1, specific for the human mAb 2F5, was inserted into 17 different positions of the glycoprotein gp64. Propagation of viruses was successful in 13 cases, mutagenesis at four positions did not result in production of intact virus particles. Western blotting, FACS analysis and ELISA were used for characterization of the different binding properties of the mutants. Insertion of this peptide into the native envelope protein resulted in high avidity display on the surface of baculovirus particles. This approach offers the possibility of effective modification of surface properties in regard to host range specificity and antigen display.
authors.affiliation: Institute of Applied Microbiology, University of Agricultural Sciences, Vienna, Austria.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Spenger
authors.forename: Reingard
authors.initials: R
authors.lastname: Grabherr
authors.forename: Lars
authors.initials: L
authors.lastname: Töllner
authors.forename: Hermann
authors.initials: H
authors.lastname: Katinger
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Ernst
mesh_terms: D000595:Amino Acid Sequence; D016367:Baculoviridae / Q000235:genetics / Q000502:physiology*; D000939:Epitopes; D015700:HIV Envelope Protein gp41 / Q000235:genetics / Q000276:immunology / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013499:Surface Properties; D014760:Viral Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D014779:Virus Replication
chemical_list: D000939:Epitopes; D015700:HIV Envelope Protein gp41; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins