CDNA encoding the human alpha2 delta4 calcium channel subunit
AssigneeOrtho-McNeil Pharmaceutical, Inc.
InventorsNing Qin, Ellen Codd
Life of this patent
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Abstract
The present invention provides nucleic acid and polypeptide sequences describing a novel isoform of the α2δ subunit of a voltage gated calcium channel, herein named as α2δ-4. The isolated nucleic acid or polypeptide molecule of the invention can be used in diagnosing and treating a disease or disorder associated with a defective α2δ-4 subunit, such as seizure-related syndromes, anxiety, multiple sclerosis, and the like.
Claims37
An isolated and purified nucleic acid molecule selected from the group consisting of:
- (a) a nucleic acid molecule comprising a nucleotide sequence having at least a 70% identity to nucleotides 1 to 224 of SEQ ID NO:9;
- (b) a nucleic acid molecule comprising a nucleotide sequence having at least a 70% identity to nucleotides 3308 to 3486 of SEQ ID NO:9;
- (c) a nucleic acid molecule comprising at least 15 sequential bases of the polynucleotide of (a) or (b);
- (d) a nucleic acid molecule comprising a nucleotide sequence that is complementary to the polynucleotide of (a), (b), or (c).
A substantially purified polypeptide selected from the group consisting of:
- (a) a polypeptide comprising an amino acid sequence having at least a 85% identity to amino acids 1 to 12 of SEQ ID NO:10;
- (b) a polypeptide comprising an amino acid sequence having at least a 70% identity to amino acids 1040 to 1090 of SEQ ID NO:10;
- and
- (c) a polypeptide comprising an amino acid sequence having at least a 85% identity to amino acids 1 to 12 of SEQ ID NO:10 and an amino acid sequence having at least 70% identity to amino acids 1040 to 1090 of SEQ ID NO:10;
- wherein the polypeptide is capable of forming a functional calcium channel.
A method for expressing an α2δ-4 calcium channel subunit protein in a recombinant host cell, comprising the steps of:
- (a) introducing an expression vector capable of encoding an α2δ-4 calcium channel subunit protein into a cell;
- and
- (b) culturing the cells under conditions that allow expression of the α2δ-4 calcium channel subunit protein from the expression vector.
An antibody that selectively binds to a peptide corresponding to amino acids 1 to 12 of SEQ ID NO:10 or amino acids 1040 to 1090 of SEQ ID NO:10.
A method of identifying variations in α2δ-4 calcium channel subunit sequences or α2δ-4 calcium channel subunit activity in a subject comprising the steps of:
- (a) obtaining a biological sample from a subject;
- (b) assaying the amount of α2δ-4 nucleic acid, α2δ-4 polypeptide, variations in the α2δ-4 nucleic acid or amino acid sequence or measuring the ionic flux through the calcium channel;
- and
- (c) comparing results from the assaying step with results using sample from a control subject.
A method of treating or preventing a disease or disorder associated with a defective α2δ-4 calcium channel subunit in a subject in need thereof, comprising the step of introducing a DNA molecule capable of encoding an α2δ-4 calcium channel subunit protein into a cell in the subject.
A method of treating or preventing a disease or disorder associated with defective α2δ-4 calcium channel subunit in a subject in need thereof, comprising the step of introducing a DNA molecule that is the antisense strand of a coding sequence capable of encoding an α2δ-4 calcium channel subunit protein into a cell in the subject.
A method of detecting a nucleic acid molecule of an α2δ-4 gene, comprising the step of contacting a test sample with a nucleic acid probe that hybridizes to the nucleic acid molecule under stringent conditions and detecting the probe-nucleic acid molecule complex.
A method of detecting a α2δ-4 calcium channel subunit protein, comprising the step of contacting a test sample with an antibody that selectively binds to amino acids 1 to 12 or 1040 to 1090 of SEQ ID NO:10;
- and detecting the protein-antibody complex.
A method of identifying a compound useful for treating a disease or disorder associated with a α2δ-4 calcium channel subunit defect, comprising the steps of:
- (a) contacting a test compound with a α2δ-4 calcium channel subunit, wherein;
- and
- (b) identifying compounds capable of increasing or decreasing the flux of ions through a calcium channel, said channel comprising the α2δ-4 calcium channel subunit.
A method for identifying a compound that alters α2δ-4 calcium channel subunit protein activity, comprising the steps of:
- (a) combining a compound, a measurably labeled ligand for the α2δ-4 calcium channel subunit protein, and a α2δ-4 calcium channel subunit protein;
- and
- (b) measuring binding of the compound to the subunit protein by a reduction in the amount labeled ligand binding to the α2δ-4 calcium channel subunit protein.
A method of identifying a compound useful for increasing or decreasing expression of an α2δ-4 calcium channel subunit protein, comprising the steps of:
- (a) contacting a test compound with a nucleic acid sequence comprising at least one regulatory sequence for an α2δ-4 gene and a reporter gene operably linked thereto;
- and
- (b) determining whether the test compound increases or decreases the expression of the reporter gene in the cell.
Dependencies are inferred from the claim text.