Stepwise transformation of a cholera toxin and a p24 (HIV-1) epitope into D-peptide analogs

Protein Eng. 1998 Oct;11(10):941-8. doi: 10.1093/protein/11.10.941.

Abstract

We have transformed two peptide epitopes into D-peptide analogs: VPGSQHIDS derived from cholera toxin recognized by the antibody TE33, and GATPQDLNTML from the HIV-1 capsid protein p24 recognized by the antibody CB4-1. The transformation process was performed by stepwise substitution of each single epitope position by all 19 D-amino acids and glycine followed by antibody binding studies and selection of one D-analog for further transformation. Thus, each transformation step introduced one novel D-position into the peptide. For both epitopes complete D-analogs were obtained. The cholera toxin-derived variant dwGsqhydp binds to the antibody TE33 with higher affinity than its original epitope, whereas in the case of the p24-derived analog saGdwwGkssl lower affinity was detected. Both D-peptides are completely stable in serum for several days. Antibody interaction models for both D-molecules were generated by computer-assisted modelling based on the crystal structures of the starting complexes. Compared with the L-peptides, the binding conformation of dwGsqhydp is very similar, whereas saGdwwGkssl displays a completely different interaction mode.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution*
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / immunology
  • Antibody Affinity
  • Cholera Toxin / immunology*
  • Computer Simulation
  • Crystallization
  • Endopeptidases / metabolism
  • Epitopes / chemistry
  • Epitopes / immunology*
  • HIV Core Protein p24 / immunology*
  • Half-Life
  • Hydrogen Bonding
  • Models, Molecular
  • Peptides / chemistry
  • Peptides / immunology*
  • Protein Conformation
  • Thermodynamics

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • HIV Core Protein p24
  • Peptides
  • Cholera Toxin
  • Endopeptidases