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PMID: 10823881 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Characterization of stable, soluble trimers containing complete ectodomains of human immunodeficiency virus type 1 envelope glycoproteins.

Journal of virology ·Vol. 74 ·No. 12 ·2000-06-00 ·Pages 5716-25

Yang X, Farzan M, Wyatt R, Sodroski J

Abstract

The human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins function as a membrane-anchored trimer of three gp120 exterior glycoproteins and three gp41 transmembrane glycoproteins. Previously, we reported three approaches to stabilize soluble trimers containing parts of the gp41 ectodomains: addition of GCN4 trimeric helices, disruption of the cleavage site between gp120 and gp41, and introduction of cysteines in the gp41 coiled coil to form intersubunit disulfide bonds. Here, we applied similar approaches to stabilize soluble gp140 trimers including the complete gp120 and gp41 ectodomains. A combination of fusion with the GCN4 trimeric sequences and disruption of the gp120-gp41 cleavage site resulted in relatively homogeneous gp140 trimers with exceptional stability. The gp120 epitopes recognized by neutralizing antibodies are intact and exposed on these gp140 trimers. By contrast, the nonneutralizing antibody epitopes on the gp120 subunits of the soluble trimers are relatively occluded compared with those on monomeric gp120 preparations. This antigenic similarity to the functional HIV-1 envelope glycoproteins and the presence of the complete gp41 ectodomain should make the soluble gp140 trimers useful tools for structural and immunologic studies.

MeSH Terms
Amino Acid Sequence Antibodies, Monoclonal/immunology Antibodies, Viral/immunology Cell Line DNA-Binding Proteins Disulfides/metabolism Epitopes/immunology Fungal Proteins/chemistry,genetics,metabolism HIV Envelope Protein gp120/chemistry,genetics,immunology,metabolism HIV Envelope Protein gp41/chemistry,genetics,immunology,metabolism HIV-1/chemistry,genetics Humans Molecular Sequence Data Molecular Weight Protein Binding Protein Conformation Protein Engineering Protein Kinases/chemistry,genetics,metabolism Protein Processing, Post-Translational Receptors, HIV/metabolism Recombinant Fusion Proteins/chemistry,genetics,metabolism Saccharomyces cerevisiae Proteins Sequence Deletion/genetics Solubility Thermodynamics
Chemicals
Antibodies, Monoclonal Antibodies, Viral DNA-Binding Proteins Disulfides Epitopes Fungal Proteins HIV Envelope Protein gp120 HIV Envelope Protein gp41 Receptors, HIV Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Protein Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Yang X
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
Farzan M
Wyatt R
Sodroski J
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.12.5716-5725.2000
Published
2000-06-00
Pages
5716-25
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC112061
Subset
IM
Grants
NIAID NIH HHS · R37 AI024755 · United States
NIAID NIH HHS · P30 AI028691 · United States
NIAID NIH HHS · AI31783 · United States
NIAID NIH HHS · R01 AI031783 · United States
NIAID NIH HHS · AI39420 · United States
NIAID NIH HHS · R01 AI039420 · United States
NIAID NIH HHS · AI24755 · United States
External Links
PubMed source DOI: 10.1128/jvi.74.12.5716-5725.2000 PMC Full Text View journal details
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