title: Expression and immunogenicity of the extracellular domain of the human immunodeficiency virus type 1 envelope glycoprotein, gp160.
abstract: The envelope glycoprotein of human immunodeficiency virus type 1 is synthesized as a precursor, gp160, that subsequently is cleaved to yield mature gp120 and gp41. In these studies, the gene encoding gp160 was mutagenized so as direct the synthesis of a truncated protein consisting of the extracellular domains of both gp120 and gp41. The variant protein, termed sgp160, consisted of 458 amino acids of gp120 and 172 amino acids of gp41. To facilitate protein purification, the normal polyglycoprotein processing site between gp120 and gp41 was deleted through the use of site-directed mutagenesis. This allowed for the synthesis of a molecule that could be purified by affinity chromatography, using acid elution, without dissociation of the gp120 polypeptide from the gp41 polypeptide. The conformation of the sgp160 variant appeared to be functionally relevant, as reflected by its ability to bind to CD4 with an affinity comparable to that of the variant rgp120. The structure of the sgp160-containing polypeptide differed from that of rgp120 in that it tended to form high-molecular-weight aggregates that could be dissociated to monomers and dimers in the presence of reducing agents. Antibodies against the sgp160 protein reacted with authentic virus-derived gp160, gp120, and gp41; neutralized viral infectivity; and inhibited the binding of rgp120 to CD4. Rabbit antibodies to the sgp160 protein differed from those raised against rgp120 in that they were enriched for populations that blocked CD4 binding but did not prevent human immunodeficiency virus type 1-induced syncytium formation.
authors.affiliation: Department of Developmental Biology, Genentech, Inc., South San Francisco, California 94080.
authors.forename: P W
authors.initials: PW
authors.lastname: Berman
authors.forename: L
authors.initials: L
authors.lastname: Riddle
authors.forename: G
authors.initials: G
authors.lastname: Nakamura
authors.forename: O K
authors.initials: OK
authors.lastname: Haffar
authors.forename: W M
authors.initials: WM
authors.lastname: Nunes
authors.forename: P
authors.initials: P
authors.lastname: Skehel
authors.forename: R
authors.initials: R
authors.lastname: Byrn
authors.forename: J
authors.initials: J
authors.lastname: Groopman
authors.forename: T
authors.initials: T
authors.lastname: Matthews
authors.forename: T
authors.initials: T
authors.lastname: Gregory
mesh_terms: D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology; D001667:Binding, Competitive; D002460:Cell Line; D002846:Chromatography, Affinity; D005786:Gene Expression Regulation; D015488:HIV Antigens / Q000276:immunology; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D009154:Mutation; D010957:Plasmids; D011233:Precipitin Tests; D012191:Retroviridae Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000945:Antigens, Differentiation, T-Lymphocyte; D015488:HIV Antigens; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins