The Carbohydrate Epitope of the Neutralizing Anti-HIV-1 Antibody 2G12

  • Christopher N. Scanlan
  • Ralph Pantophlet
  • Mark R. Wormald
  • Erica Ollmann Saphire
  • Daniel Calarese
  • Robyn Stanfield
  • Ian A. Wilson
  • Hermann Katinger
  • Raymond A. Dwek
  • Dennis R. Burton
  • Pauline M. Rudd
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 535)

Abstract

2G12 is a broadly neutralizing human monoclonal antibody against human immunodeficiency virus type-1 (HIV-1) that has previously been shown to bind to a carbohydrate-dependent epitope on gp120. Here, site-directed mutagenesis and carbohydrate analysis were used to define further the 2G12 epitope. Alanine scanning mutagenesis showed that elimination of the N-linked carbohydrate attachment sequences associated with residues N295, N332, N339, N386, and N392 by N → A substitution produced significant decreases in 2G12 binding affinity to gp120JR.CSF. The mutagenesis studies provided no convincing evidence for the involvement of gp120 amino acid side chains in 2G12 binding. Antibody binding was inhibited when gp120 was treated with Aspergillus saitoi mannosidase, Jack Bean mannosidase, or endoglycosidase H, indicating that Manal → 2Man-linked sugars of oligomannose glycans on gp120 are required for 2G12 binding. Consistent with this finding, the binding of 2G12 to gp120 could be inhibited by monomeric mannose but not by other hexoses. The data presented here suggests that the most likely epitope for 2G12 is formed from a specific cluster of mannose residues on the outer face of gp120, with the other glycans playing an indirect role in maintaining epitope conformation. [Journal of Virology, July 2002, pp. 7306–7321, Vol. 76, No. 14.]

Keywords

Carbohydrate Influenza Alanine Maned Fructose 

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Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Christopher N. Scanlan
    • 1
    • 2
  • Ralph Pantophlet
    • 2
  • Mark R. Wormald
    • 1
  • Erica Ollmann Saphire
    • 2
    • 3
  • Daniel Calarese
    • 3
  • Robyn Stanfield
    • 3
  • Ian A. Wilson
    • 3
    • 4
  • Hermann Katinger
    • 5
  • Raymond A. Dwek
    • 1
  • Dennis R. Burton
    • 2
  • Pauline M. Rudd
    • 1
  1. 1.The Glycobiology Institute, Department of BiochemistryUniversity of OxfordOxfordUK
  2. 2.Department of ImmunologyThe Scripps Research InstituteLa JollaUSA
  3. 3.Department of Molecular BiologyThe Scripps Research InstituteLa JollaUSA
  4. 4.Skaggs Institute for Chemical Biology The Scripps Research InstituteLa JollaUSA
  5. 5.Institute of Applied Microbiology, University of AgricultureViennaAustria

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