Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Minireviews
    • JVI Classic Spotlights
    • Archive
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JVI
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Journal of Virology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Minireviews
    • JVI Classic Spotlights
    • Archive
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JVI
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Structure and Assembly

Modifications That Stabilize Human Immunodeficiency Virus Envelope Glycoprotein Trimers in Solution

Xinzhen Yang, Lauren Florin, Michael Farzan, Peter Kolchinsky, Peter D. Kwong, Joseph Sodroski, Richard Wyatt
Xinzhen Yang
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute,
Department of Pathology and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lauren Florin
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael Farzan
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute,
Department of Pathology and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Peter Kolchinsky
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Peter D. Kwong
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Joseph Sodroski
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute,
Department of Pathology and
Department of Immunology and Infectious Diseases, Harvard School of Public Health,Boston, Massachusetts 02115, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard Wyatt
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute,
Department of Medicine, Harvard Medical School, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/JVI.74.10.4746-4754.2000
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Fig. 1.
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    Structure of the soluble HIV-1 envelope glycoprotein variants. The partial structures of the soluble gp120, gp140, and gp130 envelope glycoproteins used in this paper are shown beneath the corresponding segment of the wild-type gp160 glycoprotein. The junction of gp120 and gp41, the positions of the N-terminal (N36) and C-terminal (C34) gp41 α-helices, and the transmembrane (TM) region are shown. The DP178 peptide used in this study contains sequences corresponding to those of the C34 helix. The positions of the serine substitutions for arginine 508 and arginine 511 are shown (S S). The position of the Cys-Cys-Gly substitution in the N-terminal helical segment (residues 576 to 578) is shown (CCG). The position of the GCN4 trimeric peptide is also shown.

  • Fig. 2.
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Characterization of the soluble HIV-1 envelope glycoprotein variants. (A) Sucrose density gradient fractions containing the soluble HIV-1 envelope glycoproteins are shown. Fraction 1 was collected from the bottom of the gradient; fraction 10 represents the top of the gradient. Fractions were precipitated by a mixture of sera from HIV-1-infected individuals. Precipitates were analyzed on SDS-polyacrylamide gels under non-reducing conditions, except for the samples [gp130(−/CCG/GCN4) + β-ME] shown in the bottom right panel, which were treated with 1.5% β-ME before gel analysis. Asterisks indicate the expected position of a gp120 monomer in each gel. (B) Radiolabeled envelope glycoproteins were precipitated from transfected cell supernatants by a mixture of sera from HIV-1-infected individuals and were analyzed under nonreducing conditions (in the absence of β-ME). (C) Precipitates of radiolabeled envelope glycoproteins were formed as for panel B and were analyzed on SDS-polyacrylamide gels after treatment with either 1.5% β-ME for 3 min at 100°C or 5% β-ME for 10 min at 100°C.

  • Fig. 3.
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    Molecular exclusion chromatography of soluble envelope glycoproteins. The YU2 gp120 and gp130(−/GCN4) glycoproteins were analyzed on a Superdex 200 column and compared with molecular size standards, the positions of which are indicated by arrows. Elution times are also shown. The aggregate peak for the gp130(−/GCN4) glycoprotein elutes before the highest molecular size standard used (660 kDa).

  • Fig. 4.
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    Recognition of the soluble HIV-1 envelope glycoprotein variants by the DP178 gp41 peptide. Radiolabeled envelope glycoproteins in transfected cell supernatants were normalized for the amount of envelope protein and then precipitated either with a mixture of sera from HIV-1-infected patients (patient sera) or with a mixture of the 1D4 antibody and the PK-C299 peptide (DP178). The PK-C299 peptide contains the residues of the DP178 peptide, which corresponds to the C-terminal gp41 helical region (C34 in Fig. 1), and the residues of the C9 peptide, which are recognized by the 1D4 antibody.

  • Fig. 5.
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    Recognition of the soluble HIV-1 envelope glycoprotein variants by monoclonal antibodies. Immunoprecipitation of the radiolabeled envelope glycoproteins was performed with a mixture of sera from HIV-1-infected individuals (patient sera) or with monoclonal antibodies. The precipitates were analyzed on SDS-polyacrylamide gels under reducing conditions. (A) The envelope glycoproteins were precipitated by a panel of anti-gp120 monoclonal antibodies that recognize discontinuous gp120 epitopes. The 39F antibody recognizes the gp120 V3 variable loop, the F91 antibody recognizes the gp120 CD4 binding site, and the 17b antibody recognizes a CD4-induced gp120 epitope. (B) The envelope glycoproteins were precipitated by monoclonal antibodies against epitopes in the C1 and C5 regions of gp120: a discontinuous epitope involving the C1 and C5 regions (C11), and linear gp120 epitopes within the first (C1) conserved region (4D4#85 and 135/9) or within the fifth (C5) conserved region (670-30D and M91).

  • Fig. 6.
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    CCR5 binding of the soluble HIV-1 envelope glycoproteins. Equivalent amounts of radiolabeled soluble envelope glycoproteins were incubated with Cf2Th/synCCR5 cells in the absence or presence of sCD4. The bound proteins were precipitated and analyzed on SDS-polyacrylamide gels, the autoradiographs of which are shown.

PreviousNext
Back to top
Download PDF
Citation Tools
Modifications That Stabilize Human Immunodeficiency Virus Envelope Glycoprotein Trimers in Solution
Xinzhen Yang, Lauren Florin, Michael Farzan, Peter Kolchinsky, Peter D. Kwong, Joseph Sodroski, Richard Wyatt
Journal of Virology May 2000, 74 (10) 4746-4754; DOI: 10.1128/JVI.74.10.4746-4754.2000

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Journal of Virology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Modifications That Stabilize Human Immunodeficiency Virus Envelope Glycoprotein Trimers in Solution
(Your Name) has forwarded a page to you from Journal of Virology
(Your Name) thought you would be interested in this article in Journal of Virology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Modifications That Stabilize Human Immunodeficiency Virus Envelope Glycoprotein Trimers in Solution
Xinzhen Yang, Lauren Florin, Michael Farzan, Peter Kolchinsky, Peter D. Kwong, Joseph Sodroski, Richard Wyatt
Journal of Virology May 2000, 74 (10) 4746-4754; DOI: 10.1128/JVI.74.10.4746-4754.2000
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About JVI
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #Jvirology

@ASMicrobiology

       

 

JVI in collaboration with

American Society for Virology

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0022-538X; Online ISSN: 1098-5514