title: The mannose-dependent epitope for neutralizing antibody 2G12 on human immunodeficiency virus type 1 glycoprotein gp120.
abstract: We have analyzed the unique epitope for the broadly neutralizing human monoclonal antibody (MAb) 2G12 on the gp120 surface glycoprotein of human immunodeficiency virus type 1 (HIV-1). Sequence analysis, focusing on the conservation of relevant residues across multiple HIV-1 isolates, refined the epitope that was defined previously by substitutional mutagenesis (A. Trkola, M. Purtscher, T. Muster, C. Ballaun, A. Buchacher, N. Sullivan, K. Srinivasan, J. Sodroski, J. P. Moore, and H. Katinger, J. Virol. 70:1100-1108, 1996). In a biochemical study, we digested recombinant gp120 with various glycosidase enzymes of known specificities and showed that the 2G12 epitope is lost when gp120 is treated with mannosidases. Computational analyses were used to position the epitope in the context of the virion-associated envelope glycoprotein complex, to determine the variability of the surrounding surface, and to calculate the surface accessibility of possible glycan- and polypeptide-epitope components. Together, these analyses suggest that the 2G12 epitope is centered on the high-mannose and/or hybrid glycans of residues 295, 332, and 392, with peripheral glycans from 386 and 448 on either flank. The epitope is mannose dependent and composed primarily of carbohydrate, with probably no direct involvement of the gp120 polypeptide surface. It resides on a face orthogonal to the CD4 binding face, on a surface proximal to, but distinct from, that implicated in coreceptor binding. Its conservation amidst an otherwise highly variable gp120 surface suggests a functional role for the 2G12 binding site, perhaps related to the mannose-dependent attachment of HIV-1 to DC-SIGN or related lectins that facilitate virus entry into susceptible target cells.
authors.affiliation: Dept. of Microbiology and Immunology, Weill Medical College of Cornell University, New York, NY 10021, USA.
authors.forename: Rogier W
authors.initials: RW
authors.lastname: Sanders
authors.forename: Miro
authors.initials: M
authors.lastname: Venturi
authors.forename: Linnea
authors.initials: L
authors.lastname: Schiffner
authors.forename: Roopa
authors.initials: R
authors.lastname: Kalyanaraman
authors.forename: Hermann
authors.initials: H
authors.lastname: Katinger
authors.forename: Kenneth O
authors.initials: KO
authors.lastname: Lloyd
authors.forename: Peter D
authors.initials: PD
authors.lastname: Kwong
authors.forename: John P
authors.initials: JP
authors.lastname: Moore
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D002240:Carbohydrate Sequence; D018604:Epitope Mapping; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D015483:HIV Antibodies / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D008358:Mannose / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D017422:Sequence Analysis, DNA
chemical_list: D000911:Antibodies, Monoclonal; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D006026:Glycoside Hydrolases; D008358:Mannose