Structural and kinetic studies of the Fab fragment of a monoclonal anti-spin label antibody by nuclear magnetic resonance

Thomas P. Theriault, Daniel J. Leahy, Michael Levitt, Harden M. McConnell, Gordon S. Rule

Research output: Contribution to journalArticlepeer-review

19 Scopus citations


Nuclear magnetic resonance has been used to study the structure of the anti-spin label antibody AN02 combining site and kinetic rates for the hapten-antibody reaction. The association reaction for the hapten dinitrophenyl-diglycine (DNP-diGly) is diffusion-limited. The activation enthalpy for association, 5·1 kcal/mol, is close to the activation enthalpy for diffusion in water. Several reliable resonance assignments have been made with the aid of recently reported crystal structure. Structural data deduced from the nuclear magnetic resonance (n.m.r.) spectra compare favorably with the crystal structure in terms of the combining site amino acid composition, distances of tyrosine residues from unpaired electron of the hapten, and residues in direct contact with the hapten. Evidence is presented that a single binding site region tyrosine residue can assume two distinct conformations on binding of DNP-diGly. The AN02 antibody is an autoantibody. Dimerization of the Fab fragments is blocked by the hapten DNP-diGly. The n.m.r. spectra suggests that some of the amino acid residues involved in the binding of the DNP-hapten are also involved in the Fab dimerization.

Original languageEnglish (US)
Pages (from-to)257-270
Number of pages14
JournalJournal of molecular biology
Issue number1
StatePublished - Sep 5 1991
Externally publishedYes


  • anti-DNP
  • autoantibody
  • monoclonal antibody
  • n.m.r.
  • nitroxide spin label

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Structural Biology


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