Screening method, a composition comprising substances chosen in the method thereof, and a binding substance

Abstract

(Problems) The main object of the present invention is to provide a screening method for selecting a substance affecting a bond between thioredoxin and MIF. (Means for Solving Problems) The present invention provides a screening method for selecting a test substance which strengthens a bond between a polypeptide of a thioredoxin family and a macrophage migration inhibition factor, comprising: mixing a test substance with at least one binding substance selected from following (1) to (4), (1) the polypeptide belonging to the thioredoxin family, (2) a protein having an amino acid sequence of the polypeptide belonging to the thioredoxin family in which one or more amino acid is deleted, replaced or added, and having an equivalent activity to the polypeptide of the thioredoxin family, (3) a gene coding (1), (4) a gene coding (2); bonding the binding substance to the macrophage migration inhibition factor; and monitoring the bond state between the binding substance and the macrophage migration inhibition factor.

1 . A screening method for selecting a test substance which strengthens a bond between a polypeptide of a thioredoxin family and a macrophage migration inhibition factor, comprising:
 mixing a test substance with at least one binding substance selected from following (1) to (4),   (1) the polypeptide belonging to the thioredoxin family   (2) a protein having an amino acid sequence of the polypeptide belonging to the thioredoxin family in which one or more amino acid is deleted, replaced or added, and having an equivalent activity to the polypeptide of the thioredoxin family   (3) a gene coding (1)   (4) a gene coding (2);   bonding the binding substance to the macrophage migration inhibition factor; and   monitoring the bond state between the binding substance and the macrophage migration inhibition factor.   
     
     
         2 . The screening method according to the  claim 1 , wherein the monitor of the bond state between the binding substance and the macrophage migration inhibition factor is performed with a molecular interaction analysis. 
     
     
         3 . A composition for strengthening a bond between a polypeptide of a thioredoxin family and a macrophage migration inhibition factor, comprising at least one selected from nitrosoglutathione or prostaglandin I2. 
     
     
         4 . A binding substance which bonds directly with a migration inhibition factor, comprising at least one selected from following (1) to (4).
 (1) a polypeptide belonging to a thioredoxin family   (2) a protein having an amino acid sequence of the polypeptide belonging to the thioredoxin family in which one or more amino acid is deleted, replaced or added, and having an equivalent activity to the polypeptide of the thioredoxin family   (3) a gene coding (1)   (4) a gene coding (2)   
     
     
         5 . The binding substance according to the  claim 4 , wherein the binding substance is a polypeptides having any one of -Cys-Gly-Pro-Cys-, -Cys-Pro-Tyr-Cys-, -Cys-Pro-His-Cys- or -Cys-Pro-Pro-Cys- in an active center.