Inhibition of the HIV-1 spike by single-PG9/16-Antibody binding suggests a coordinated-activation model for its three protomeric units

Robin Löving, Mathilda Sjöberg, Shang-Rung Wu, James M. Binley, Henrik Garoff

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The HIV-1 spike is composed of three protomeric units, each containing a peripheral gp120 and a transmembrane gp41 subunit. Binding to the CD4 and the chemokine receptors triggers them to mediate virus entry into cells by membrane fusion. The spikes also represent the major target for neutralizing antibodies (Abs) against the virus. We have studied how two related broadly neutralizing Abs, PG9 and PG16, react with the spike. Unexpectedly, this also suggested how the functions of the individual protomers in the spike depend on each other. The Abs have been shown to bind the V1/V2 loops of gp120, located at the top of the spike. Using blue native-polyacrylamide gel electrophoresis (BN-PAGE), we show that only single Abs or antigen-binding fragments could bind to the spikes of HIV-1 virus-like particles. Apparently, binding to one gp120 sterically interferes with binding to the other two subunits in the spike top. Despite this constraint, all of the protomers of the spike became resistant to CD4 binding and subsequent formation of the coreceptor binding site. These activities were measured by monitoring the sequential complex formation of the spike first with Abs and then with soluble 2d- or 4d-CD4 or with soluble CD4 and the CD4 inducible coreceptor binding site Ab 17b in BN-PAGE. The inhibition of the spike by single-Ab binding suggested that the activation reactions of the individual protomeric units are linked to each other in a coordinated activation process.

Original languageEnglish
Pages (from-to)7000-7007
Number of pages8
JournalJournal of Virology
Volume87
Issue number12
DOIs
Publication statusPublished - 2013 Jun 1

Fingerprint

Human immunodeficiency virus 1
Native Polyacrylamide Gel Electrophoresis
HIV-1
Protein Subunits
protein subunits
Neutralizing Antibodies
neutralizing antibodies
antibodies
polyacrylamide gel electrophoresis
Antibodies
binding sites
Binding Sites
Virus Internalization
viruses
CD4 Antigens
Membrane Fusion
virus-like particles
Cell Fusion
Chemokine Receptors
Virion

All Science Journal Classification (ASJC) codes

  • Immunology
  • Virology

Cite this

Löving, Robin ; Sjöberg, Mathilda ; Wu, Shang-Rung ; Binley, James M. ; Garoff, Henrik. / Inhibition of the HIV-1 spike by single-PG9/16-Antibody binding suggests a coordinated-activation model for its three protomeric units. In: Journal of Virology. 2013 ; Vol. 87, No. 12. pp. 7000-7007.
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Inhibition of the HIV-1 spike by single-PG9/16-Antibody binding suggests a coordinated-activation model for its three protomeric units. / Löving, Robin; Sjöberg, Mathilda; Wu, Shang-Rung; Binley, James M.; Garoff, Henrik.

In: Journal of Virology, Vol. 87, No. 12, 01.06.2013, p. 7000-7007.

Research output: Contribution to journalArticle

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