Patent ID: 6322972
Filing Date: 2001-11-27
Classification: A61K,C07K

Abstract:
A method of identifying an inhibitor of TNF-R death domain binding which comprises:(a) transforming cells with a first polynucleotide encoding a TNF-R death domain protein, a second polynucleotide encoding a TNF-R1-DD ligand protein, and at least one reporter gene, wherein the expression of the reporter gene is regulated by the binding of the TNF-R1-DD ligand protein encoded by the second polynucleotide to the TNF-R death domain protein encoded by the first polynucleotide and wherein said second polynucleotide is selected from the group consisting of:(i) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:1 from nucleotide 2 to nucleotide 1231;(ii) a polynucleotide comprising a fragment of nucleotide sequence of SEQ ID NO:1, which encodes a protein having TNF-R-DD ligand protein activity;(iii) a polynucleotide encoding an TNF-R-DD ligand protein comprising the amino acid sequence of SEQ ID NO:2;(iv) a polynucleotide encoding an TNF-R1-DD ligand protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 and having TNF-R1-DD ligand protein activity;(v) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 2 to nucleotide 415;(vi) a polynucleotide comprising a fragment of the nucleotide of SEQ ID NO:3, which encodes a protein having TNF-R1-DD ligand protein activity;(vii) a polynucleotide encoding an TNF-R1-DD ligand protein comprising the amino acid sequence of SEQ ID NO:4;(viii) a polynucleotide encoding an TNF-R1 -DD ligand protein comprising a fragment of the amino acid sequence of SEQ ID NO:4 and having TNF-R1 -DD ligand protein activity;(ix) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 from nucleotide 2 to nucleotide 559;(x) a polynucleotide comprising a fragment of the nucleotide sequence of SEQ ID NO:5, which encodes a protein having TNF-R1-DD ligand protein activity;(xi) a polynucleotide encoding an TNF-R1-DD ligand protein comprising the amino acid sequence of SEQ ID NO:6;(xii) a polynucleotide encoding an TNF-R1-DD ligand protein comprising a fragment of the amino acid sequence of SEQ ID NO:6 and having TNF-R1-DD ligand protein activity;(xiii) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7 from nucleotide 57 to nucleotide 875;(ix) a polynucleotide comprising a fragment of the nucleotide sequence of SEQ ID NO:7, which encodes a protein having TNF-R1-DD ligand protein activity;(xv) a polynucleotide encoding an TNF-RI -DD ligand protein comprising the amino acid sequence of SEQ ID NO:8;(xvi) a polynucleotide encoding an TNF-R1-DD ligand protein comprising a fragment of the amino acid sequence of SEQ ID NO:8 and having TNF-R1-DD ligand protein activity; and(xvii) a polynucleotide which encodes a protein having a TNF-R1-DD ligand protein activity and hybridizes in 6.times.SSC at 65 degrees C. to a polynucleotide which is complementary to any one of the polynucleotides specified in (i)-(xvi);(b) growing transformed cells in the presence of and in the absence of a compound; and(c) comparing the degree of expression of the reporter gene in the presence of and in the absence of the compound,wherein the compound is capable of inhibiting TNF-R death domain binding when a decrease in the degree of expression of the reporter gene occurs.