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Identification of an N-linked glycan in the V1-loop of HIV-1 gp120 influencing neutralization by anti-V3 antibodies and soluble CD4

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Summary

Glycosylation is necessary for HIV-1 gp120 to attain a functional conformation, and individual N-linked glycans of gp120 are important, but not essential, for replication of HIV-1 in cell culture. We have constructed a mutant HIV-1 infectious clone lacking a signal for N-linked glycosylation in the V1-loop of HIV-1 gp120. Lack of an N-linked glycan was verified by a mobility enhancement of mutant gp120 in SDS-gel electrophoresis. The mutated virus showed no differences in either gp120 content per infectious unit or infectivity, indicating that the N-linked glycan was neither essential nor affecting viral infectivity in cell culture. We found that the mutated virus lacking an N-linked glycan in the V1-loop of gp120 was more resistant to neutralization by monoclonal antibodies to the V3-loop and neutralization by soluble recombinant CD4 (sCD4). Both viruses were equally well neutralized by ConA and a conformation dependent human antibody IAM-2G12. This suggests that the N-linked glycan in the V1-loop modulates the three-dimensional conformation of gp120, without changing the overall functional integrity of the molecule.

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Authors and Affiliations

  1. Laboratory for Infectious Diseases, Hvidovre Hospital, Hvidovre, Denmark

    G. J. Gram, S. Olofsson & J. E. S. Hansen

  2. Department of Clinical Virology, University of Göteborg, Göteborg, Sweden

    A. Hemming, A. Bolmstedt, B. Jansson, S. Olofsson & J. O. Nielsen

  3. Biomedical Center, Department of Virology, The National Veterinary Institute, Uppsala, Sweden

    L. Åkerblom

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  1. G. J. Gram
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  2. A. Hemming
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  3. A. Bolmstedt
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  4. B. Jansson
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  5. S. Olofsson
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  6. L. Åkerblom
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  7. J. O. Nielsen
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  8. J. E. S. Hansen
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Gram, G.J., Hemming, A., Bolmstedt, A. et al. Identification of an N-linked glycan in the V1-loop of HIV-1 gp120 influencing neutralization by anti-V3 antibodies and soluble CD4. Archives of Virology 139, 253–261 (1994). https://doi.org/10.1007/BF01310789

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  • DOI: https://doi.org/10.1007/BF01310789

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