CARD3X-2 polypeptides, encoding nucleic acids, and methods of use
The invention provides a polypeptide corresponding to a newly identified splice variant of the CARD3X gene, termed CARD3X-2. Also provided by the invention is an encoding nucleic acid molecule, as well as a CARD3X-2 specific antibody. The invention further provides related screening, diagnostic and therapeutic methods.
1 . An isolated nucleic acid molecule encoding a CARD3X-2 polypeptide comprising the amino acid sequence set forth as SEQ ID NO:197, or a domain of said polypeptide selected from a CARD domain, NACHT domain, and LRR domain.
2 . The nucleic acid molecule of claim 1 , which encodes a CARD3X-2 polypeptide comprising the amino acid sequence set forth as SEQ ID NO:197.
3 . The nucleic acid molecule of claim 1 , wherein the nucleotide sequence of said nucleic acid molecule comprises SEQ ID NO:196.
4 . The nucleic acid molecule of claim 1 , wherein said nucleic acid molecule is cDNA.
5 . A vector containing the nucleic acid molecule of claim 1 .
6 . Recombinant cells containing the nucleic acid molecule of claim 1 .
7 . An oligonucleotide comprising at least 15 contiguous nucleotides of the nucleic acid molecule of claim 3 , or the complement thereof.
8 . An oligonucleotide according to claim 7 , wherein said oligonucleotide is labeled with a detectable marker.
9 . An isolated CARD3X-2 polypeptide, comprising the amino acid sequence set forth as SEQ ID NO:197, or a domain of said polypeptide selected from a CARD domain, NACHT domain, and LRR domain.
10 . The isolated CARD3X-2 polypeptide of claim 9 , comprising the amino acid sequence set forth as SEQ ID NO:197.
11 . A method of producing a CARD3X-2 polypeptide comprising expressing the cDNA of claim 4 in vitro or in a cell under conditions suitable for expression of said polypeptide.
12 . An isolated anti-CARD3X-2 antibody having specific reactivity with the CARD3X-2 polypeptide of claim 9 .
13 . The antibody of claim 12 , wherein said antibody is a monoclonal antibody.
14 . A cell line producing the monoclonal antibody of claim 13 .
15 . The antibody of claim 12 , wherein said antibody is a polyclonal antibody.
16 . A method for detecting the presence of a CARD3X-2 polypeptide in a sample, comprising contacting a test sample with an antibody according to claim 12 or a recombinant phage, detecting the presence of an antibody:CARD3X-2 complex or recombinant phage:CARD3X-2 complex, and thereby detecting the presence of a CARD3X-2 polypeptide in said sample.
17 . A method of identifying a CARD3X-2 binding molecule comprising:
(a) contacting the CARD3X-2 polypeptide of claim 9 with a candidate CARD3X-2 binding molecule;
(b) detecting association of CARD3X-2 polypeptide with said CARD3X-2 binding molecule.
18 . The method of claim 17 , wherein the CARD3X-2 binding molecule is a CARD3X-2-associated polypeptide.
19 . The method of claim 17 , wherein the CARD3X-2 binding molecule is a small molecule.
20 . A method of identifying an effective agent that alters association of a NACHT-containing polypeptide with a NACHT-associated polypeptide (NAP), comprising the steps of:
(a) contacting a NACHT-containing polypeptide selected from SEQ ID NOS:188, 189 and 197, and said NAP with an agent suspected of being able to alter the association of said NACHT-containing polypeptide and said NAP, under conditions that allow association between said NACHT-containing polypeptide and said NAP; and
(b) detecting the altered association of said NACHT-containing polypeptide and said NAP, wherein said altered association identifies an effective agent.
21 . The method of claim 20 , wherein said NAP is selected from CARD3X, CARD3X-2, Nod1, NAC, PAN2, NAIP and cyropyrin.
22 . A CARD3X-2 mRNA targeting molecule, comprising a molecule selected from an anti-sense oligonucleotide, a ribozyme and an si RNA, wherein said molecule binds selectively to an mRNA corresponding to the nucleotide sequence referenced as SEQ ID NO:196, or a portion of said nucleotide sequence.