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

  • Conference paper
Glycobiology and Medicine

Part of the book series: Advances in Experimental Medicine and Biology ((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.]

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Authors and Affiliations

  1. The Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK

    Christopher N. Scanlan, Mark R. Wormald, Raymond A. Dwek & Pauline M. Rudd

  2. Department of Immunology, The Scripps Research Institute, La Jolla, California, 92037, USA

    Christopher N. Scanlan, Ralph Pantophlet, Erica Ollmann Saphire & Dennis R. Burton

  3. Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, 92037, USA

    Erica Ollmann Saphire, Daniel Calarese, Robyn Stanfield & Ian A. Wilson

  4. Skaggs Institute for Chemical Biology The Scripps Research Institute, La Jolla, California, 92037, USA

    Ian A. Wilson

  5. Institute of Applied Microbiology, University of Agriculture, 1190, Vienna, Austria

    Hermann Katinger

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  1. Christopher N. Scanlan
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  2. Ralph Pantophlet
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  7. Ian A. Wilson
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  1. The Sir Joseph Hotung Centre for Musculoskeletal Diseases St Georges Hospital and Medical School, London, UK

    John S. Axford (Director) (Director)

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Scanlan, C.N. et al. (2003). The Carbohydrate Epitope of the Neutralizing Anti-HIV-1 Antibody 2G12. In: Axford, J.S. (eds) Glycobiology and Medicine. Advances in Experimental Medicine and Biology, vol 535. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0065-0_13

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  • DOI: https://doi.org/10.1007/978-1-4615-0065-0_13

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