Expression and antigenicity of human immunodeficiency virus type-1 transmembrane protein GP41 in insect cells
Journal
Biochemical and Biophysical Research Communications
Journal Volume
191
Journal Issue
1
Pages
207-213
Date Issued
1993
Author(s)
Abstract
The HIV-1 transmembrane protein, gp41, is processed together with the envelope glycoprotein, gp120, from the same precursor, gp160, during the virus maturation. We used a baculovirus expression system to demonstrate that gp41 could be properly expressed without the preceding gp120 sequence. Two constructs with slight differences in the N-terminal region of gp41 were generated: one with a deletion of the first 7 hydrophobic residues of gp41, which have been suggested to be in a region important for membrane fusion and penetration, whereas the second with a complete sequence of gp41 except that a nonconserved leucine was substituted with a glutamine during DNA manipulation. Results from Western blotting with specific antisera confirm the gp41 identity. The sizes of gp41 were sensitive to tunicamycin treatment, indicating that N-linked glycosylation did occur. Further immunoblotting analyses with 90 different serum samples from HIV-1 -infected individuals gave similar reaction patterns, suggesting that gp120 as well as the N-terminal region of gp4l are not critical for the expression and antigenecity of gp41. These eucaryotic constructs should provide valuable gp41 sources for detailed characterization of gp41 functions. ? 1993 Academic Press.
SDGs
Other Subjects
glycoprotein; membrane protein; amino acid sequence; animal cell; article; cell membrane; gene expression; human immunodeficiency virus; immunoblotting; insect; nonhuman; nucleotide sequence; priority journal; Amino Acid Sequence; Animal; Baculoviridae; Base Sequence; Blotting, Western; Cell Line; Epitopes; Gene Expression; Genetic Vectors; HIV Envelope Protein gp120; HIV Envelope Protein gp41; HIV-1; Insects; Molecular Sequence Data; Mutagenesis, Insertional; Oligodeoxyribonucleotides; Plasmids; Restriction Mapping; Support, Non-U.S. Gov't; Transfection; Tunicamycin
Type
journal article