Structural analysis of chick ephrin-A2 by function-blocking and non-blocking monoclonal antibodies

Biochem Biophys Res Commun. 2002 Jul 12;295(2):348-53. doi: 10.1016/s0006-291x(02)00681-2.

Abstract

Ephrins, ligands for the Eph family of receptor tyrosine kinases, play key roles in diverse biological processes. In this study, we determined the epitopes and kinetic parameters of function-blocking (B3) and non-blocking (IV) monoclonal antibodies (mAbs) recognizing chick ephrin-A2. We show that the epitope for the non-blocking mAb is the residue Asp(105) of chick ephrin-A2. However, the binding of the function-blocking mAb depends mostly on residue Ser(108) and its epitope may reside within residues 105-132, which appear crucial for the receptor interaction site. Kinetic studies suggest a possible mechanism why mAb IV, despite recognizing a region very close to the mAb B3 epitope, fails to block the ligand-receptor interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Base Sequence
  • Cell Line
  • Chickens
  • DNA Primers
  • Ephrin-A2
  • Humans
  • Kinetics
  • Ligands
  • Protein Conformation
  • Recombinant Fusion Proteins / immunology
  • Surface Plasmon Resonance
  • Transcription Factors / antagonists & inhibitors
  • Transcription Factors / chemistry*

Substances

  • Antibodies, Monoclonal
  • DNA Primers
  • Ephrin-A2
  • Ligands
  • Recombinant Fusion Proteins
  • Transcription Factors