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PMID: 12719580 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Molecular features of the broadly neutralizing immunoglobulin G1 b12 required for recognition of human immunodeficiency virus type 1 gp120.

Journal of virology ·Vol. 77 ·No. 10 ·2003-05-00 ·Pages 5863-76

Zwick MB, Parren PW, Saphire EO, Church S, Wang M, Scott JK, Dawson PE, Wilson IA, Burton DR

Abstract

IgG1 b12 is a broadly neutralizing antibody against human immunodeficiency virus type 1 (HIV-1). The epitope recognized by b12 overlaps the CD4 receptor-binding site (CD4bs) on gp120 and has been a target for vaccine design. Determination of the three-dimensional structure of immunoglobulin G1 (IgG1) b12 allowed modeling of the b12-gp120 interaction in which the protruding third complementarity-determining region (CDR) of the heavy chain (H3) was crucial for antibody binding. In the present study, extensive mutational analysis of the antigen-binding site of Fab b12 was carried out to investigate the validity of the model and to identify residues important for gp120 recognition and, by inference, key to the anti-HIV-1 activity of IgG1 b12. In all, 50 mutations were tested: 40 in H3, 4 each in H2 and L1, and 2 in L3. The results suggest that the interaction of gp120 with H3 of b12 is crucially dependent not only on a Trp residue at the apex of the H3 loop but also on a number of residues at the base of the loop. The arrangement of these residues, including aromatic side chains and side chains that hydrogen bond across the base of the loop, may rigidify H3 for penetration of the recessed CD4-binding cavity. The results further emphasize the importance to gp120 binding of a Tyr residue at the apex of the H2 loop that forms a second finger-like structure and a number of Arg residues in L1 that form a positively charged, shelf-like structure. In general, the data are consistent with the b12-gp120 interaction model previously proposed. At the gene level, somatic mutation is seen to be crucial for the generation of many of the structural features described. The Fab b12 mutants were also tested against the b12 epitope-mimic peptide B2.1, and the reactivity profile had many similarities but also significant differences from that observed for gp120. The paratope map of b12 may facilitate the design of molecules that are able to elicit b12-like activities.

MeSH Terms
Amino Acid Sequence Binding Sites Complementarity Determining Regions/chemistry,genetics HIV Antibodies/chemistry,genetics,immunology HIV Envelope Protein gp120/immunology HIV-1/immunology Humans Immunoglobulin Fab Fragments/chemistry,genetics,immunology Immunoglobulin G/chemistry,genetics,immunology Immunoglobulin Heavy Chains/chemistry,genetics,immunology Models, Molecular Molecular Sequence Data Mutagenesis Neutralization Tests Sequence Analysis, DNA
Chemicals
Complementarity Determining Regions HIV Antibodies HIV Envelope Protein gp120 Immunoglobulin Fab Fragments Immunoglobulin G Immunoglobulin Heavy Chains
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Zwick Michael B
Department of Immunology, The Scripps Research Institute, La Jolla, California 92037, USA.
Parren Paul W H I
Saphire Erica O
Church Sarah
Wang Meng
Scott Jamie K
Dawson Philip E
Wilson Ian A
Burton Dennis R
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.77.10.5863-5876.2003
Published
2003-05-00
Pages
5863-76
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC154005
Subset
IM
Grants
NIGMS NIH HHS · R01 GM046192 · United States
NIGMS NIH HHS · GM46192 · United States
NIAID NIH HHS · R01 AI049111 · United States
NIMH NIH HHS · P30 MH062261 · United States
NIAID NIH HHS · AI49808 · United States
NIAID NIH HHS · R01 AI049111-01A1 · United States
NIAID NIH HHS · R37 AI033292 · United States
NIAID NIH HHS · AI49111 · United States
NIAID NIH HHS · R01 AI033292 · United States
NIAID NIH HHS · AI43077 · United States
NIAID NIH HHS · R21 AI049808-02 · United States
NIAID NIH HHS · AI33292 · United States
NIMH NIH HHS · MH62261 · United States
Databases
GENBANK
UNKNOWN
External Links
PubMed source DOI: 10.1128/jvi.77.10.5863-5876.2003 PMC Full Text View journal details
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