Home LiteratureArticle Details
PMID: 16174734 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Dissection of the carbohydrate specificity of the broadly neutralizing anti-HIV-1 antibody 2G12.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 102 ·No. 38 ·2005-09-20 ·Pages 13372-7

Calarese DA, Lee HK, Huang CY, Best MD, Astronomo RD, Stanfield RL, Katinger H, Burton DR, Wong CH, Wilson IA

Abstract

Human antibody 2G12 neutralizes a broad range of HIV-1 isolates. Hence, molecular characterization of its epitope, which corresponds to a conserved cluster of oligomannoses on the viral envelope glycoprotein gp120, is a high priority in HIV vaccine design. A prior crystal structure of 2G12 in complex with Man(9)GlcNAc(2) highlighted the central importance of the D1 arm in antibody binding. To characterize the specificity of 2G12 more precisely, we performed solution-phase ELISA, carbohydrate microarray analysis, and cocrystallized Fab 2G12 with four different oligomannose derivatives (Man(4), Man(5), Man(7), and Man(8)) that compete with gp120 for binding to 2G12. Our combined studies reveal that 2G12 is capable of binding both the D1 and D3 arms of the Man(9)GlcNAc(2) moiety, which would provide more flexibility to make the required multivalent interactions between the antibody and the gp120 oligomannose cluster than thought previously. These results have important consequences for the design of immunogens to elicit 2G12-like neutralizing antibodies as a component of an HIV vaccine.

MeSH Terms
AIDS Vaccines/immunology Antibodies, Monoclonal/chemistry,immunology Binding Sites, Antibody/immunology Carbohydrate Sequence Crystallography, X-Ray Epitopes/chemistry,immunology HIV Antibodies/chemistry,immunology HIV Envelope Protein gp120/immunology HIV-1/chemistry,immunology Mannose/chemistry,immunology Molecular Sequence Data Oligosaccharides, Branched-Chain/chemistry,immunology
Chemicals
AIDS Vaccines Antibodies, Monoclonal Epitopes HIV Antibodies HIV Envelope Protein gp120 Oligosaccharides, Branched-Chain Mannose
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Calarese Daniel A
Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Lee Hing-Ken
Huang Cheng-Yuan
Best Michael D
Astronomo Rena D
Stanfield Robyn L
Katinger Hermann
Burton Dennis R
Wong Chi-Huey
Wilson Ian A
References (34)
34 references, click to expand
  1. Further additions to MolScript version 1.4, including reading and contouring of electron-density maps.
    Acta Crystallogr D Biol Crystallogr. 1999 Apr;55(Pt 4):938-40 PMID: 10089341
  2. Synthesis of the Globo H Hexasaccharide Using the Programmable Reactivity-Based One-Pot Strategy This research was supported by the National Institutes of Health. F.B. thanks the Deutsche Forschungsgemeinschaft for a fellowship.
    Angew Chem Int Ed Engl. 2001 Apr 1;40(7):1274-1277 PMID: 11301448
  3. The mannose-dependent epitope for neutralizing antibody 2G12 on human immunodeficiency virus type 1 glycoprotein gp120.
    J Virol. 2002 Jul;76(14):7293-305 PMID: 12072528
  4. The broadly neutralizing anti-human immunodeficiency virus type 1 antibody 2G12 recognizes a cluster of alpha1-->2 mannose residues on the outer face of gp120.
    J Virol. 2002 Jul;76(14):7306-21 PMID: 12072529
  5. Saccharide display on microtiter plates.
    Chem Biol. 2002 Jun;9(6):713-20 PMID: 12079783
  6. The chemistry of amine-azide interconversion: catalytic diazotransfer and regioselective azide reduction.
    J Am Chem Soc. 2002 Sep 11;124(36):10773-8 PMID: 12207533
  7. Reactivity-based one-pot synthesis of a Lewis Y carbohydrate hapten: a colon-rectal cancer antigen determinant.
    Angew Chem Int Ed Engl. 2002 Nov 4;41(21):4087-90 PMID: 12412090
  8. Synthesis of sugar arrays in microtiter plate.
    J Am Chem Soc. 2002 Dec 4;124(48):14397-402 PMID: 12452714
  9. Reactivity-based one-pot total synthesis of fucose GM1 oligosaccharide: a sialylated antigenic epitope of small-cell lung cancer.
    Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):797-802 PMID: 12552090
  10. A new reactivity-based one-pot synthesis of N-acetyllactosamine oligomers.
    J Org Chem. 2003 Mar 21;68(6):2135-42 PMID: 12636372
  11. Synthesis of monoglucosylated high-mannose-type dodecasaccharide, a putative ligand for molecular chaperone, calnexin, and calreticulin.
    J Am Chem Soc. 2003 Mar 26;125(12):3402-3 PMID: 12643681
  12. Antibody domain exchange is an immunological solution to carbohydrate cluster recognition.
    Science. 2003 Jun 27;300(5628):2065-71 PMID: 12829775
  13. Immunopathogenesis and immunotherapy in AIDS virus infections.
    Nat Med. 2003 Jul;9(7):861-6 PMID: 12835706
  14. HIV vaccines 1983-2003.
    Nat Med. 2003 Jul;9(7):874-80 PMID: 12835708
  15. Design and synthesis of a template-assembled oligomannose cluster as an epitope mimic for human HIV-neutralizing antibody 2G12.
    Org Biomol Chem. 2004 Feb 21;2(4):483-8 PMID: 14770226
  16. Reactivity-based one-pot synthesis of oligomannoses: defining antigens recognized by 2G12, a broadly neutralizing anti-HIV-1 antibody.
    Angew Chem Int Ed Engl. 2004 Feb 13;43(8):1000-3 PMID: 14966891
  17. HIV vaccine design and the neutralizing antibody problem.
    Nat Immunol. 2004 Mar;5(3):233-6 PMID: 14985706
  18. Likelihood-enhanced fast rotation functions.
    Acta Crystallogr D Biol Crystallogr. 2004 Mar;60(Pt 3):432-8 PMID: 14993666
  19. Probing protein-carbohydrate interactions with microarrays of synthetic oligosaccharides.
    Chembiochem. 2004 Mar 5;5(3):379-82 PMID: 14997532
  20. Binding of high-mannose-type oligosaccharides and synthetic oligomannose clusters to human antibody 2G12: implications for HIV-1 vaccine design.
    Chem Biol. 2004 Jan;11(1):127-34 PMID: 15113002
  21. In pursuit of carbohydrate-based HIV vaccines, part 1: The total synthesis of hybrid-type gp120 fragments.
    Angew Chem Int Ed Engl. 2004 May 3;43(19):2557-61 PMID: 15127452
  22. In pursuit of carbohydrate-based HIV vaccines, part 2: The total synthesis of high-mannose-type gp120 fragments--evaluation of strategies directed to maximal convergence.
    Angew Chem Int Ed Engl. 2004 May 3;43(19):2562-5 PMID: 15127453
  23. Oligosaccharide and glycoprotein microarrays as tools in HIV glycobiology; glycan-dependent gp120/protein interactions.
    Chem Biol. 2004 Jun;11(6):875-81 PMID: 15217620
  24. Covalent display of oligosaccharide arrays in microtiter plates.
    J Am Chem Soc. 2004 Jul 21;126(28):8640-1 PMID: 15250702
  25. Toward fully synthetic carbohydrate-based HIV antigen design: on the critical role of bivalency.
    J Am Chem Soc. 2004 Aug 11;126(31):9560-2 PMID: 15291558
  26. The CCP4 suite: programs for protein crystallography.
    Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3 PMID: 15299374
  27. Printed covalent glycan array for ligand profiling of diverse glycan binding proteins.
    Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):17033-8 PMID: 15563589
  28. Proteins that bind high-mannose sugars of the HIV envelope.
    Prog Biophys Mol Biol. 2005 Jun;88(2):233-82 PMID: 15572157
  29. Raster3D: photorealistic molecular graphics.
    Methods Enzymol. 1997;277:505-24 PMID: 18488322
  30. Processing of X-ray diffraction data collected in oscillation mode.
    Methods Enzymol. 1997;276:307-26 PMID: 27754618
  31. Structure of myohemerythrin in the azidomet state at 1.7/1.3 A resolution.
    J Mol Biol. 1987 Sep 20;197(2):273-96 PMID: 3681996
  32. Diffraction methods for biological macromolecules. Interactive computer graphics: FRODO.
    Methods Enzymol. 1985;115:157-71 PMID: 3841179
  33. The molecular surface package.
    J Mol Graph. 1993 Jun;11(2):139-41 PMID: 8347567
  34. Crystallography & NMR system: A new software suite for macromolecular structure determination.
    Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21 PMID: 9757107
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0505763102
Published
2005-09-20
Epub
2005-00-07
Pages
13372-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1224641
Subset
IM
Grants
NIGMS NIH HHS · R01 GM046192 · United States
NIGMS NIH HHS · GM46192 · United States
NIAID NIH HHS · R37 AI033292 · United States
NIAID NIH HHS · R01 AI033292 · United States
NIAID NIH HHS · AI33292 · United States
Databases
PDB
1ZLS, 1ZLU, 1ZLV, 1ZLW
External Links
PubMed source DOI: 10.1073/pnas.0505763102 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2