Screening method
View Patent ↗The present invention intends to provide a screening of agonist/antagonist and the like. Specifically, the invention provides a method of screening a compound or its salt that alters the binding property between a novel G protein-coupled receptor protein or its salt, and a substance relating to cholesterol metabolism, which comprises using the receptor protein or its salt, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 14 or SEQ ID NO: 16, and the substance relating to cholesterol metabolism.
1. A method of screening a compound or salts thereof that alters the binding property between a G protein-coupled receptor protein or salts thereof, and a bile acid, wherein said method comprises:
a) contacting a labeled bile acid with the G protein-coupled receptor protein containing the amino acid sequence represented by SEQ ID NO: 1 or a protein containing an amino acid sequence having at least 90% homology to the amino acid sequence represented by SEQ NO: 1 and having the same ligand binding activity as the amino acid sequence represented by SEQ ID NO: 1 or salts thereof, and measuring the amount of said labeled bile acid bound to said G protein-coupled receptor protein or said protein or salts thereof; and
b) contacting a labeled bile acid and a test compound with the G protein-coupled receptor protein containing the amino acid sequence represented by SEQ ID NO: 1 or a protein containing an amino acid sequence having at least 90% homology to the amino acid sequence represented by SEQ NO: 1 and having the same ligand binding activity as the amino acid sequence represented by SEQ ID NO: 1 or salts thereof, and measuring the amount of said labeled bile acid, bound to said G protein-coupled or said protein or salts thereof; and
c) comparing binding property between a) and b).
2. A kit for screening a compound or salts thereof that alters the binding property between a G protein-coupled receptor protein or salts thereof, and a bile acid, which comprises (1) the G protein-coupled receptor protein containing the amino acid sequence represented by SEQ ID NO: 1 or a protein containing an amino acid sequence having at least 90% homology to the amino acid sequence represented by SEQ NO: 1 and having the same ligand binding activity as the amino acid sequence represented by SEQ ID NO: 1, or salts thereof, and (2) the bile acid.