Methods of screening for antagonists of mammalian monocyte chemoattractant protein receptors
View Patent ↗Novel human chemokine receptors, MCP-1RA and MCP-1RB, and processes for producing them are disclosed. The receptors, which are alternately spliced versions of MCP-1 receptor protein may be used in an assay to identify antagonists of MCP-1 which are therapeutically useful in the treatment of atherosclerosis and other diseases characterized by monocytic infiltrates.
1. A method to identify an antagonist of MCP-1R, comprising:
(a) contacting an isolated MCP-1R polypeptide with a candidate antagonist, wherein the MCP-1R polypeptide comprises an amino acid sequence at least 95% homologous to an extracellular domain of SEQ ID NO: 2 or of SEQ ID NO: 4, and
(b) comparing the binding of MCP-1 to the MCP-1R polypeptide in the presence of the candidate antagonist to the binding of MCP-1 to the MCP-1R polypeptide in its absence;
wherein a candidate antagonist which causes a decrease in the binding of MCP-1 to the MCP-1R polypeptide is identified as an antagonist of MCP-1R.
2. The method of claim 1 , wherein the MCP-1R polypeptide is a MCP-1RA polypeptide.
3. The method of claim 2 , wherein the MCP-1RA polypeptide comprises SEQ ID NO: 2.
4. The method of claim 1 , wherein the MCP-1R polypeptide is a MCP-1RB polypeptide.
5. The method of claim 4 wherein the MCP-1RB polypeptide comprises SEQ ID NO: 4.
6. The method of claim 1 , wherein the MCP-1 is detectably labeled.
7. The method of claim 1 , further comprising the step of introducing an expression vector encoding the MCP-1R polypeptide into a cell, wherein the MCP-1R polypeptide is expressed on the surface of a cell.
8. The method of claim 1 , wherein the MCP-1R polypeptide consists essentially of an MCP-1R extracellular domain.
9. The method of claim 1 , wherein the MCP-1R polypeptide is a chimeric protein.