Structural characterization of mouse monoclonal antibody 13-1 against a porphyrin derivative: identification of a disulfide bond in CDR-H3 of Mab 13-1

Biochem Biophys Res Commun. 1997 Nov 26;240(3):566-72. doi: 10.1006/bbrc.1997.7668.

Abstract

The amino acid sequence of a mouse monoclonal antibody Mab13-1, a catalytic antibody against TCPP (meso-tetrakis(4-carboxyphenyl)porphyrin), was confirmed by mass spectrometric (MS) peptide mapping. The amino-terminal sequence of the heavy chain was established by MS/MS analysis of the isolated N-terminal peptide. The presence of a unique disulfide bond between Cys93H and Cys102H was identified by MS peptide mapping and sequence analysis of an S-S containing peptide. Positions of other disulfide bonds were identified to be conserved. The non-conserved disulfide bridge was found to be resistant as other intra-chain disulfide bonds against reduction under non-denaturing condition, and to be buried inside the molecule. This extra disulfide bond is expected to support antigen-binding by restricting the flexibility of CDR-H3 loop, and it might be favorable for the recognition of a plane antigen, a porphyrin derivative.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Catalytic / chemistry*
  • Antibodies, Catalytic / immunology
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal, Murine-Derived
  • Cysteine / chemistry
  • Disulfides / chemistry*
  • Immunoglobulin Variable Region / chemistry*
  • Immunoglobulin Variable Region / immunology
  • Mice
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Peptide Mapping
  • Porphyrins / immunology*
  • Sequence Analysis
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Antibodies, Catalytic
  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Murine-Derived
  • Disulfides
  • Immunoglobulin Variable Region
  • Mab13-1 monoclonal antibody
  • Peptide Fragments
  • Porphyrins
  • tetracarboxyphenylporphine
  • Cysteine