Significant structural and functional change of an antigen-binding site by a distant amino acid substitution: proposal of a structural mechanism

Proc Natl Acad Sci U S A. 1989 Jul;86(14):5532-6. doi: 10.1073/pnas.86.14.5532.

Abstract

To study the molecular basis for antibody diversity and the structural basis for antigen binding, we have characterized the loss of phosphocholine (P-Cho) binding both experimentally and computationally in U10, a somatic mutant of the antibody S107. Nucleotide sequencing of U10 shows a single base change in JH1, substituting Asp-101 with Ala, over 9 A distant from the P-Cho-binding pocket. Probing with antiidiotypic antibodies suggests local, not global, conformational changes. Computational results support a specific structural mechanism for the loss of P-Cho binding. The U10 mutation eliminates the charged interaction between Asp-101 and Arg-94, which allows the Arg-94 side chain to disrupt P-Cho binding sterically and electrostatically by folding into the P-Cho-binding site. These results specifically show the importance of the Arg-94 to Asp-101 side chain salt bridge in the heavy-chain CDR3 conformation and suggest that residues distant from the binding site play an important role in antibody diversity and inducible complementarity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies / genetics*
  • Antibody Diversity
  • Antigen-Antibody Complex*
  • Antigens*
  • Base Sequence
  • Binding Sites
  • Cell Line
  • Computer Simulation
  • Enzyme-Linked Immunosorbent Assay
  • Gene Rearrangement*
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation
  • Phosphorylcholine / immunology
  • Protein Conformation
  • Structure-Activity Relationship

Substances

  • Antibodies
  • Antigen-Antibody Complex
  • Antigens
  • Phosphorylcholine

Associated data

  • GENBANK/M24452