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Related Experiment Videos

Computer analysis of mutations that affect antibody specificity.

J Novotny1, R E Bruccoleri, E Haber

  • 1Massachusetts General Hospital, Boston 02114.

Proteins
|January 1, 1990
PubMed
Summary
This summary is machine-generated.

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Antibody specificity for cardiac glycosides like digoxin was altered by mutations distant from the binding site. Structural analysis revealed how these mutations, particularly at heavy chain position 94, influence antibody-antigen interactions and affinity.

Area of Science:

  • Immunology
  • Structural Biology
  • Computational Chemistry

Background:

  • The 40-150 hybridoma cell line produces high-affinity antibodies against cardiac glycosides (digoxin, digitoxin).
  • Mutant cell lines 40-150 A2.4 and 40-150 A2.4 P.10 exhibit altered antibody specificities.
  • Key mutations occur at heavy chain position 94 (Ser to Arg) and N-terminal residue deletions.

Purpose of the Study:

  • To elucidate the structural basis for altered antibody specificity in mutants 40-150 A2.4 and 40-150 A2.4 P.10.
  • To understand how mutations distant from the antigen-binding site affect antibody-antigen interactions.

Main Methods:

  • Computer-based structural modeling of the 40-150 antibody and its mutants, using McPC 603 coordinates.
  • Conformational analysis using the CONGEN program to sample torsional space around position H94.

Related Experiment Videos

  • Detailed analysis of lowest energy conformations to identify structural changes and potential interactions.

    • Main Results:

    • Substitution of Ser-H94 with Arg-H94 allows hydrogen bonding with Asp-H101, Arg-L46, and Asp-L55, altering the combining site surface.
    • This alteration in the combining site surface is proposed to explain changes in antibody affinity.
    • Deletion of N-terminal residues in mutant 40-150 A2.4 P.10 increases Arg-H94 solvent accessibility, potentially disrupting the Arg-H94-Asp-H101 hydrogen bond and restoring original specificity.

    Conclusions:

    • Mutations distant from the antibody's antigen-binding site can significantly impact specificity and affinity through allosteric effects.
    • Hydrogen bonding networks involving Arg-H94 play a crucial role in modulating antibody-antigen interactions.
    • Solvation effects can further influence these interactions, demonstrating the complex interplay of structural factors in antibody function.