Structural properties of HIV-1 Env fused with the 14-kDa vaccinia virus envelope protein.
To gain insights into the structure-function relationship of the envelope (env) glycoprotein of the human immunodeficiency virus type 1 (HIV-1) we have generated a vaccinia virus (VV) recombinant (VV-14kENV) that expresses a fusion protein (14k-env) consisting of the VV 14-kDa envelope protein (110 amino acids) fused at the C-terminus with HIV-1 env protein (816 amino acids). The 14k-env protein displayed unique structural properties in virus-infected cells. This protein was recognized by 14 kDa-specific antisera as well as HIV-1 env antisera. It was not cleaved during virus infection of cultured cells of various origins, it was stable, it was not released to the medium, and it was not incorporated into virions. Instead of a predicted 174-kDa protein, two proteins of about 110 and 100 kDa were observed. The size reduction of the fusion protein was due to limited glycosylation (110 kDa) and formation of unglycosylated protein (100 kDa). The 14k-env protein formed oligomeric structures and was exposed on the cell surface after virus infection. When mice were inoculated with the recombinant virus that expresses the 14K-env fusion protein, humoral immune response against gp160 was observed. Our findings suggest that 14k-env protein might display novel immunogenic properties.
Rodriguez
J R
JR
Department of Biochemistry, State University of New York, Brooklyn 11203.
Rodriguez
D
D
Esteban
M
M
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D002478:Cells, Cultured; D015686:Gene Products, env / Q000276:immunology / Q000378:metabolism*; D006031:Glycosylation; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000378:metabolism*; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D008970:Molecular Weight; D011498:Protein Precursors / Q000276:immunology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D014616:Vaccinia virus / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure
D000914:Antibodies, Viral; D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D011994:Recombinant Proteins