Receptor-ligand system and assay
View Patent ↗An Eph family RTK (receptor tyrosine kinase) ligand-binding domain as well as a method of identifying Eph family RTK agonists or antagonist. The ligand-binding domain is suitably encoded by exon III of a gone encoding the RTK of the Eph family and may additionally include an amino acid sequence encoded by exon II of the gene. The ligand-binding domain may be a polypeptide having amino acids 57-271 of the sequence shown in FIG. 1.
1. An isolated polypeptide which binds a ligand selected from the group consisting of LERK3, LERK4, LERK5 and LERK7, wherein said isolated polypeptide consists of the amino acid sequence set forth in SEQ ID NO:4.
2. The isolated polypeptide of claim 1 , wherein said polypeptide has at least one pair of disulfide cysteine residues selected from the group consisting of:
(i) Cys 71 -Cys 189 ; and
(ii) Cys 259 -Cys 270 ,
wherein said cysteine residues are identified in SEQ ID NO:4.
3. An isolated polypeptide which binds a LERK, said polypeptide consisting of the amino acid sequence of SEQ ID NO:4.
4. The polypeptide of claim 3 , wherein the LERK is LERK7.
5. The isolated polypeptide of claim 1 which is a recombinant polypeptide produced by a host cell.
6. A method of identifying a molecule which binds the polypeptide of claim 1 , which method includes the steps of:
(i) combining a sample suspected of containing said molecule with the polypeptide of claim 1 ; and
(ii) determining if said molecule is present in said sample by measuring the binding of the molecule to said polypeptide; wherein binding identifies said molecule.
7. A method of identifying a molecule which competes with binding of a ligand to the polypeptide of claim 1 , which method includes the steps of:
(i) combining a sample suspected of containing said molecule, a ligand and at the polypeptide of claim 1 ; and
(ii) determining if the molecule is present in the sample by measuring the competitive binding of the molecule and the ligand with said polypeptide; wherein a molecule that competes with ligand for binding to said polypeptide identifies said molecule.