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Application of NMR to Conformational Studies of an HIV Peptide Bound to a Neutralizing Antibody

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NMR in Supramolecular Chemistry

Part of the book series: NATO ASI Series ((ASIC,volume 526))

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Abstract

NMR spectroscopy has become an important tool for structural studies of proteins and macro molecular complexes. In case of an antibody-antigen complex, different approaches are taken to alleviate the size limit, which is beyond NMR capabilities. The method of choice depends on the kinetic properties of the system under investigation and on the antibody fragment available. We have recently studied the conformation of RP135, a 24-residue HIV-1 peptide corresponding to the principal neutralizing determinant of the virus envelop glycoprotein gp120, in complex with 0.5β, a neutralizing antibody raised against gp120. The binding of the peptide to the antibody was too strong to observe TRNOE, therefore 2D-NOESY difference spectroscopy was applied using three strategies: (a) deuteration of specific residues of the peptide; (b) Arg→Lys replacement and; (c) truncation of the peptide antigen. The restraints on interproton distances within the bound peptide were used to calculate its conformation. The peptide forms a 10-residue loop, while the two segments flanking this loop interact extensively with each other and possibly form antiparallel β-strands.

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Authors and Affiliations

  1. Weizmann Institute of Science, Rehovot 76100, Israel

    Anat Zvi & Jacob Anglister

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  1. Anat Zvi
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  2. Jacob Anglister
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  1. Departament de Química Orgànica, Universitat de Barcelona, Spain

    M. Pons

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Zvi, A., Anglister, J. (1999). Application of NMR to Conformational Studies of an HIV Peptide Bound to a Neutralizing Antibody. In: Pons, M. (eds) NMR in Supramolecular Chemistry. NATO ASI Series, vol 526. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-4615-9_9

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  • DOI: https://doi.org/10.1007/978-94-011-4615-9_9

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-5950-3

  • Online ISBN: 978-94-011-4615-9

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