Novel molecules of the card-related protein family and uses thereof
AssigneeMillennium Pharmaceuticals, Inc.
InventorJohn Bertin
Life of this patent
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Abstract
Novel CARD-12 polypeptides, proteins, and nucleic acid molecules are disclosed. In addition to isolated CARD-12 proteins, the invention further provides CARD-12, fusion proteins, antigenic peptides and anti-CARD-12 antibodies. The invention also provides CARD-12 nucleic acid molecules, recombinant expression vectors containing a nucleic acid molecule of the invention, host cells into which the expression vectors have been introduced and non-human transgenic animals in which a CARD-12 gene has been introduced or disrupted. Diagnostic, screening and therapeutic methods utilizing compositions of the invention are also provided.
Claims18
A method for identifying a compound which binds to CARD-12, the method comprising the steps of:
- a) contacting a polypeptide, or a cell from the group consisting of:
- i) a polypeptide comprising an amino acid sequence at least 90% identical to SEQ ID NO:2;
- ii) a polypeptide comprising an amino acid sequence at least 90% identical to the CARD domain (amino acid residues 1-88 of SEQ ID NO:2) of CARD-12;
- iii) a polypeptide comprising an amino acid sequence at least 90% identical to an NBS domain (amino acid residues 161-323 of SEQ ID NO:2) of CARD-12;
- iv) a polypeptide comprising an amino acid sequence at least 90% identical to a LRR domain (amino acid residues 762-965 of SEQ ID NO:2) of CARD-12;
- v) a polypeptide comprising an amino acid sequence of a fragment of at least 200 amino acids of SEQ ID NO:2, wherein the fragment has at least one binding activity of CARD-12;
- and
- vi) a cell expressing the polypeptide of i), ii), iii), iv) or v), with a test compound;
- and
- b) determining whether the polypeptide binds to the test compound.
A method for identifying a candidate modulator of CARD-12, the method comprising:
- a) contacting a polypeptide selected from the group consisting of:
- i) a polypeptide comprising an amino acid sequence at least 90% identical to amino acid residues 161-323 of SEQ ID NO:2;
- ii) a polypeptide comprising an amino acid sequence at least 90% identical to amino acid residues 169-456 of SEQ ID NO:2 of CARD-12;
- and
- iii) a polypeptide with a length of at least 25 amino acids, comprising at least one NACHT NTPase domain (amino acid residues 169-186, 196-220, 229-253, 261-282, 330-351, 414-430, or 438-457 of SEQ ID NO:2) of CARD-12,
- with a test compound in the presence of a nucleotide that binds to the polypeptide in the absence of the test compound;
- b) measuring the binding of the nucleotide to the polypeptide in the presence of the test compound;
- and
- c) identifying the test compound as a candidate modulator of CARD-12 if the test compound increases or decreases the binding of the nucleotide to the polypeptide.
A method for identifying a candidate compound for treating an apoptotic or inflammatory disorder, the method comprising:
- a) measuring the binding of a first polypeptide or cell selected from the group consisting of:
- i) a polypeptide comprising an amino acid sequence at least 90% identical to SEQ ID NO:2;
- ii) a polypeptide comprising an amino acid sequence at least 90% identical to the CARD domain (amino acid residues 1-88 of SEQ ID NO:2) of CARD-12;
- iii) a polypeptide comprising an amino acid sequence at least 90% identical to amino acid residues 161-323 of SEQ ID NO:2 of CARD-12;
- iv) a polypeptide comprising an amino acid sequence at least 90% identical to amino acid residues 762-965 of SEQ ID NO:2 of CARD-12;
- v) a polypeptide comprising an amino acid sequence at least 90% identical to amino acid residues 169-456 of SEQ ID NO:2) of CARD-12;
- vi) a polypeptide with a length of at least 25 amino acids, comprising at least one NACHT NTPase domain (amino acid residues 169-186, 196-220, 229-253, 261-282, 330-351, 414-430, or 438-457 of SEQ ID NO:2) of CARD-12;
- vii) a polypeptide with a length of at least 50 amino acids, comprising at least one leucine-rich repeat domain (amino acid residues 656-686, 687-708, 711-737, 738-761, 762-788, 789-817, 819-845, 846-874, 875-901, 903-930, 936-962, 965-993, or 994-1021 of SEQ ID NO:2) of CARD-12;
- and
- viii) a cell expressing the polypeptide of i), ii), iii), iv), v) vi) or vii), to a second molecule selected from the group consisting of:
- a polypeptide comprising a CARD domain, caspase-1 and a nucleotide, in the presence of a test compound;
- and
- b) comparing the binding of the first polypeptide or cell to the second molecule measured in step (a) to the binding of the first polypeptide or cell to the second molecule in the absence of the test compound,
- wherein altered binding of the first polypeptide or cell to the second molecule in the presence of the test compound compared the binding in the absence of the test compound indicates that the test compound is candidate compound treating a disorder.
Dependencies are inferred from the claim text.