Ligand for EPH-related kinase (LERK) 7 is the preferred high affinity ligand for the HEK receptor
Details
Publication Year 1997-06-27,Volume 272,Issue #26,Page 16521-16530
Journal Title
JOURNAL OF BIOLOGICAL CHEMISTRY
Publication Type
Journal Article
Abstract
HEK is a member of the EPH-like receptor tyrosine kinase family, which appear to have roles in development and oncogenesis. Recently, we purified a soluble HEK ligand which is also a ligand (AL1) for the HEK-related receptor EHK1. Promiscuity appears to be a characteristic feature of interactions between the EPH-like receptors and their ligands, termed ligands for EPH-related kinases (LERKs). This prompted us to analyze the interactions between the HER exodomain and fusion proteins comprising candidate LERKs and the Fc portion of human IgG(1) (Fc) or a FLAG(TM)-peptide tag by surface plasmon resonance, size exclusion high performance liquid chromatography, sedimentation equilibrium, and transphosphorylation. Our results indicate that AL1/LERK7 is the preferred high-affinity ligand for HEK, forming a stable 1:1 complex with a dissociation constant of 12 nM. As expected the apparent affinities of bivalent fusion proteins of LERKs and the Fc portion of human IgG(1) had significantly reduced dissociation rates compared with their monovalent, FLAG(TM)-tagged derivatives. High-avidity binding of monovalent ligands can be achieved by antibody-mediated cross-linking of monovalent ligands and with LERK7 results in specific phosphorylation of the receptor. By extrapolation, our findings indicate that some of the reported LERK-receptor interactions are a consequence of the use of bivalent ligand or receptor constructs and may be functionally irrelevant.
Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Keywords
PROTEIN-TYROSINE KINASE; SURFACE-PLASMON RESONANCE; BINDING CONSTANTS; MONOCLONAL-ANTIBODY; MOLECULAR-CLONING; KINETIC-ANALYSIS; GROWTH-FACTOR; CELL LINES; FAMILY; EXPRESSION
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Creation Date: 1997-06-27 12:00:00
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