title: Structural analysis of chick ephrin-A2 by function-blocking and non-blocking monoclonal antibodies.
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.
authors.affiliation: Division of Developmental Neurobiology, Kumamoto University Graduate School of Medical Sciences, Honjo 2, Kumamoto 860-0811, Japan.
authors.forename: Mansoor
authors.initials: M
authors.lastname: Ahsan
authors.forename: Yanzhi
authors.initials: Y
authors.lastname: Yin
authors.forename: Takamasa
authors.initials: T
authors.lastname: Ueno
authors.forename: Masafumi
authors.initials: M
authors.lastname: Takiguchi
authors.forename: Hideaki
authors.initials: H
authors.lastname: Tanaka
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D017931:DNA Primers; D036383:Ephrin-A2; D006801:Humans; D007700:Kinetics; D008024:Ligands; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000276:immunology; D020349:Surface Plasmon Resonance; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; D036383:Ephrin-A2; D008024:Ligands; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors