Method of screening TGF-beta-inhibiting substances

Abstract

A method for screening substances that inhibit binding between a TAK1 polypeptide and a TAB1 polypeptide, which comprises contacting the TAB1 polypeptide to the TAK1 polypeptide and a test sample and then detecting or determining the TAK1 polypeptide that is bound to the TAB1 polypeptide.

1 . A method for screening substances that inhibit binding between a TAK1 polypeptide and a TAB1 polypeptide, which method comprises contacting a TAB1 polypeptide to a TAK1 polypeptide and a test sample, and then detecting or determining the TAK1 polypeptide that is bound to the TAB1 polypeptide.  
     
     
         2 . A method for screening substances that inhibit binding between the TAK1 polypeptide and the TAB1 polypeptide, which method comprises contacting a TAK1 polypeptide to a TAB1 polypeptide and a test sample, and then detecting or determining the TAB1 polypeptide that is bound to the TAK1 polypeptide.  
     
     
         3 . The screening method according to  claim 1  or  2 , wherein the TAB1 polypeptide has an amino acid sequence comprising Met at amino acid position 1 to Pro at amino acid position 504 of the amino acid sequence as set forth in SEQ ID NO: 2 or having an amino acid sequence modified by the substitution, deletion and/or addition of one or a plurality of amino acid residues of the amino acid sequence as set forth in SEQ ID NO: 2 and maintaining the biological activity of the TAB1 polypeptide.  
     
     
         4 . The screening method according to  claim 1  or  2 , wherein the TAK1 polypeptide has an amino acid sequence comprising Met at amino acid position 1 to Ser at amino acid position 579 of the amino acid sequence as set forth in SEQ ID NO: 4 or having an amino acid sequence modified by the substitution, deletion and/or addition of one or a plurality of amino acid residues of the amino acid sequence as set forth in SEQ ID NO: 4 and maintaining the biological activity of the TAK1 polypeptide.  
     
     
         5 . The screening method according to any of  claims 1  to  4 , wherein the TAB1 polypeptide is fused to another peptide or polypeptide.  
     
     
         6 . The screening method according to any of  claims 1  to  4 , wherein the TAK1 polypeptide is fused to another peptide or polypeptide.  
     
     
         7 . The screening method according to any of  claims 3  to  6 , wherein the TABI polypeptide is bound to a support.  
     
     
         8 . The screening method according to any of  claims 2  to  6 , wherein the TAK1 polypeptide is bound to a support.  
     
     
         9 . The screening method according to  claim 7  or  8  wherein the support is beads or a plate.  
     
     
         10 . The screening method according to  claims 1  to  6  wherein the TAK1 polypeptide, the TAB1 polypeptide, and the test sample are contacted under a homogeneous condition.  
     
     
         11 . The screening method according to any of  claim 1  and  claims 3  to  10 , wherein the TAK1 polypeptide is a labeled TAK1 polypeptide and said label is detected or determined.  
     
     
         12 . The screening method according to any of  claims 2  to  10 , wherein the TAB1 polypeptide is a labeled TAB1 polypeptide and said label is detected or determined.  
     
     
         13 . The screening method according to  claim 11  or  12  wherein the labeled TAK1 polypeptide or the labeled TAB1 polypeptide is labeled with a radioisotope, an enzyme or a fluorescent substance.  
     
     
         14 . The screening method according to any of  claims 3  to  10 , wherein the TAK1 polypeptide that is bound to the TAB1 polypeptide is detected or determined by a primary antibody against the TAK1 polypeptide or a primary antibody against another peptide or polypeptide that is fused to the TAK1 polypeptide.  
     
     
         15 . The screening method according to any of  claims 2  to  10 , wherein the TAK1 polypeptide that is bound to the TAB1 polypeptide is detected or determined by a primary antibody against the TAK1 polypeptide or a primary antibody against another peptide or polypeptide that is fused to the TAK1 polypeptide, and a secondary antibody against the primary antibody.  
     
     
         16 . The screening method according to any of claims  3  to  10 , wherein the TAB1 polypeptide that is bound to the TAK1 polypeptide is detected or determined by a primary antibody against the TAB1 polypeptide or a primary antibody against another peptide or polypeptide, that is fused to the TAB1 polypeptide.  
     
     
         17 . The screening method according to any of  claims 2  to  10 , wherein the TAB1 polypeptide that is bound to the TAK1 polypeptide is detected or determined by a primary antibody against the TAB1 polypeptide or a primary antibody against another peptide or polypeptide that is fused to the TAB1 polypeptide, and a secondary antibody against the primary antibody.  
     
     
         18 . The screening method according to any of  claims 14  to  17 , wherein the primary antibody or secondary antibody is labeled with a radioisotope, an enzyme or a fluorescent substance.  
     
     
         19 . A substance obtained by the screening method according to any of  claims 1  to  18 .  
     
     
         20 . A substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         21 . An inhibitor of signal transduction of TGF-β comprising a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         22 . An activator of signal transduction of TGF-β comprising a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         23 . A suppressor of the enhanced production of extracellular matrix protein comprising as. an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         24 . An activator of the enhanced production of extracellular matrix protein comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         25 . A suppressor of the inhibition of cellular growth comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         26 . An activator of the inhibition of cellular growth comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         27 . A suppressor of monocyte migration comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         28 . An activator of monocyte migration comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  8 .  
     
     
         29 . A suppressor of the induction of biologically active substances comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by the screening method according to any of  claims 1  to  18 .  
     
     
         30 . An activator of the induction of biologically active substances comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         31 . A suppressor of immunosuppressive action comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         32 . An activator of immunosuppressive action comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         33 . A suppressor of the deposition of amyloid β protein comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         34 . An activator of the deposition of amyloid β protein comprising as an active ingredient a substance that inhibits binding between a TAK1 polypeptide and a TAB1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         35 . A substance that inhibits the function of a TAK1 polypeptide obtained by a screening method according to any of  claims 1  to  18 .  
     
     
         36 . The substance according to  claim 35  in which the function is a kinase activity.