Novel TWIK-6, TWIK-7, IC23927, TWIK-8, IC47611, IC47615, HNMDA-1, TWIK-9, alpha2delta-4, 8099, 46455, 54414, 53763, 97076, 97102, 44181, 67084Fl and 67084alt molecules and uses therefor
The invention provides isolated nucleic acids molecules, designated TWIK-6, TWIK-7, IC23927, TWIK-8, IC47611, IC47615, HNMDA-1, TWIK-9, α 2 δ-4, 8099, 46455, 54414, 53763, 97076, 97102, 44181, 67084FL and 67084alt nucleic acid molecules, which encode novel ion channel family molecules, including calcium channels, potassium channels, and NMDA receptors. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing TWIK-6, TWIK-7, IC23927, TWIK-8, IC47611, IC47615, HNMDA-1, TWIK-9, α 2 δ-4, 8099, 46455, 54414, 53763, 97076, 97102, 44181, 67084FL and 67084alt nucleic acid molecules, host cells into which the expression vectors have been introduced, and non-human transgenic animals in which a TWIK-6, TWIK-7, IC23927, TWIK-8, IC47611, IC47615, HNMDA-1, TWIK-9, α 2 δ-4, 8099, 46455, 54414, 53763, 97076, 97102, 44181, 67084FL and 67084alt gene has been introduced or disrupted. The invention still further provides isolated TWIK-6, TWIK-7, IC23927, TWIK-8, IC47611, IC47615, HNMDA-1, TWIK-9, α 2 δ-4, 8099, 46455, 54414, 53763, 97076, 97102, 44181, 67084FL and 67084alt polypeptides, fusion polypeptides, antigenic peptides and anti-TWIK-6, TWIK-7, IC23927, TWIK-8, IC47611, IC47615, HNMDA-1, TWIK-9, α 2 δ-4, 8099, 46455, 54414, 53763, 97076, 97102, 44181, 67084FL and 67084alt antibodies. Diagnostic and therapeutic methods utilizing compositions of the invention are also provided.
1 . An isolated nucleic acid molecule selected from the group consisting of:
(a) a nucleic acid molecule comprising a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 14, 16, 21, 23, 28, 30, 31, 33, 34, 36, 37, 39, 40, 42, 43, 45, 46, 48, 49, 51, 52, 54, 55, 57, 58, 60, 61, 63, 64, 66, 67, 69, 70 or 72, or a complement thereof;
(b) a nucleic acid molecule comprising a fragment of at least 30 nucleotides of a nucleic acid comprising the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 14, 16, 21, 23, 28, 30, 31, 33, 34, 36, 37, 39, 40, 42, 43, 45, 46, 48, 49, 51, 52, 54, 55, 57, 58, 60, 61, 63, 64, 66, 67, 69, 70 or 72, or a complement thereof;
(c) a nucleic acid molecule which encodes a polypeptide comprising an amino acid sequence at least about 90% identical to the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71, or a complement thereof; and
(d) a nucleic acid molecule which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71, wherein the fragment comprises at least 10 contiguous amino acid residues of the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71, or a complement thereof.
2 . The isolated nucleic acid molecule of claim 1 , wherein the isolated nucleic acid is selected from the group consisting of:
(a) a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:1, 4, 14, 21, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, 67, or 70, or a complement thereof;
(b) a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:3, 6, 16, 23, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, or 72, or a complement thereof; and
(c) a nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68, 71 or a complement thereof.
3 . An isolated nucleic acid molecule comprising the nucleic acid molecule of claim 1 and a nucleotide sequence encoding a heterologous polypeptide.
4 . A vector comprising the nucleic acid molecule of claim 1 .
5 . The vector of claim 4 , which is an expression vector.
6 . A host cell transfected with the expression vector of claim 5 .
7 . A method of producing a polypeptide comprising culturing the host cell of claim 6 in an appropriate culture medium, to thereby produce the polypeptide.
8 . An isolated polypeptide selected from the group consisting of:
a) a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71;
b) a polypeptide consisting of the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71;
c) a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71, wherein the fragment comprises at least 10 contiguous amino acids of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71;
d) a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to complement of a nucleic acid molecule consisting of SEQ ID NO:1, 3, 4, 6, 14, 16, 21, 23, 28, 30, 31, 33, 34, 36, 37, 39, 40, 42, 43, 45, 46, 48, 49, 51, 52, 54, 55, 57, 58, 60, 61, 63, 64, 66, 67, 69, 70 or 72 under stringent conditions;
e) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 90% identical to a nucleic acid comprising the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 14, 16, 21, 23, 28, 30, 31, 33, 34, 36, 37, 39, 40, 42, 43, 45, 46, 48, 49, 51, 52, 54, 55, 57, 58, 60, 61, 63, 64, 66, 67, 69, 70 or 72; and
f) a polypeptide comprising an amino acid sequence which is at least 90% identical to the amino acid sequence of SEQ ID NO:2, 5, 15, 22, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68 or 71.
9 . The polypeptide of claim 8 , further comprising heterologous amino acid sequences.
10 . An antibody which selectively binds to a polypeptide of claim 8 .
11 . A method for detecting the presence of a polypeptide of claim 8 in a sample comprising:
a) contacting the sample with a compound which selectively binds to the polypeptide; and
b) determining whether the compound binds to the polypeptide in the sample to thereby detect the presence of a polypeptide of claim 8 in the sample.
12 . The method of claim 11 , wherein the compound which binds to the polypeptide is an antibody.
13 . A kit comprising a compound which selectively binds to a polypeptide of claim 8 and instructions for use.
14 . A method for detecting the presence of a nucleic acid molecule of claim 1 in a sample comprising:
a) contacting the sample with a nucleic acid probe or primer which selectively hybridizes to the nucleic acid molecule; and
b) determining whether the nucleic acid probe or primer binds to a nucleic acid molecule in the sample to thereby detect the presence of a nucleic acid molecule of claim 1 in the sample.
15 . The method of claim 14 , wherein the sample comprises mRNA molecules and is contacted with a nucleic acid probe.
16 . A kit comprising a compound which selectively hybridizes to a nucleic acid molecule of claim 1 and instructions for use.
17 . A method for identifying a compound which binds to a polypeptide of claim 8 comprising:
a) contacting the polypeptide, or a cell expressing the polypeptide with a test compound; and
b) determining whether the polypeptide binds to the test compound.
18 . The method of claim 17 , wherein the binding of the test compound to the polypeptide is detected by a method selected from the group consisting of:
a) detection of binding by direct detection of test compound/polypeptide binding;
b) detection of binding using a competition binding assay; and
c) detection of binding using an assay for TWIK-6, TWIK-7, IC23927, TWIK-8, IC47611, IC47615, HNMDA-1, TWIK-9, α 2 δ-4, 8099, 46455, 54414, 53763, 97076, 97102, 44181, 67084FL and 67084alt activity.
19 . A method for modulating the activity of a polypeptide of claim 8 comprising contacting the polypeptide or a cell expressing the polypeptide with a compound which binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide.
20 . A method for identifying a compound which modulates the activity of a polypeptide of claim 8 comprising:
a) contacting a polypeptide of claim 8 with a test compound; and
b) determining the effect of the test compound on the activity of the polypeptide to thereby identify a compound which modulates the activity of the polypeptide.