Claims
- 1. 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).
- 2. The nucleic acid molecule of claim 1 wherein the polynucleotide is RNA.
- 3. The nucleic acid molecule of claim 1 wherein the polynucleotide is DNA.
- 4. The isolated and purified nucleic acid molecule of claim 1 wherein the nucleic acid molecule comprises a nucleotide sequence having at least a 85% identity to nucleotides 1 to 224 of SEQ ID NO:9 or the complementary sequence thereof; wherein the nucleic acid molecule encodes a polypeptide that is capable of forming a functional calcium channel.
- 5. The isolated and purified nucleic acid molecule of claim 1 comprising a nucleotide sequence having at least a 85% identity to nucleotides 3308 to 3486 of SEQ ID NO:9 or the complementary sequence thereof; wherein the nucleic acid molecule encoding a polypeptide that is capable of forming a functional calcium channel.
- 6. The isolated and purified nucleic acid molecule of claim 1, wherein the nucleic acid molecule comprises a nucleotide sequence having at least a 85% identity to nucleotides 1 to 224 of SEQ ID NO:9 and a nucleotide sequence having at least a 85% identity to nucleotides 3308 to 3486 of SEQ ID NO:9 or the complementary sequence thereof; wherein the nucleic acid molecule encodes a polypeptide that is capable of forming a functional calcium channel.
- 7. The isolated and purified nucleic acid molecule of claim 1, encoding a polypeptide having at least a 96% identity to amino acids 1 to 1090 of SEQ ID NO:10, or the complementary sequence thereof.
- 8. The isolated and purified nucleic acid molecule of claim 1, encoding a polypeptide comprising amino acids 1 to 1090 of SEQ ID NO:10, or the complementary sequence thereof.
- 9. The isolated and purified nucleic acid molecule of claim 1, comprising at least a 94% identity to a polynucleotide sequence shown in SEQ ID NO:9, or the complementary sequence thereof.
- 10. The isolated and purified nucleic acid molecule of claim 1, having a nucleotide sequence of SEQ.ID.NO:9, or the complementary sequence thereof.
- 11. An expression vector comprising the nucleic acid molecule of claim 1.
- 12. A recombinant host cell comprising the expression vector of claim 11.
- 13. 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.
- 14. The substantially purified polypeptide of claim 13 comprising an amino acid sequence having at least a 85% identity to amino acids 1040 to 1090 of SEQ ID NO:10.
- 15. The substantially purified polypeptide of claim 13 comprising an amino acid sequence having identity to amino acids 1 to 12 and having an amino acid sequence with at least an 85% identity to amino acids 1040 to 1090 of SEQ ID NO:10.
- 16. The substantially purified polypeptide of claim 13 comprising an amino acid sequence having at least a 96% identity to amino acids 1 to 1090 of SEQ ID NO:10.
- 17. The substantially purified polypeptide of claim 13 comprising the amino acid sequence of: SEQ.ID.NO:10.
- 18. 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.
- 19. 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.
- 20. 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.
- 21. The method of claim 20, wherein the disease is selected from the group consisting of: seizure-related syndromes, epilepsy, migraine, ataxia, vestibular defects, chronic pain, neuropathic pain, mood, sleep interference, anxiety, ALS, multiple sclerosis, mania, tremor, parkinsonism, substance abuse/addiction syndromes, depression, cancer, and inflammation.
- 22. 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.
- 23. 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.
- 24. 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.
- 25. 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.
- 26. 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.
- 27. The method of claim 26 wherein said α2δ-4 calcium channel subunit is isolated or substantially purified.
- 28. The method of claim 26 wherein said α2δ-4 calcium channel subunit is associated with a membrane.
- 29. The method of claim 26 wherein said α2δ-4 calcium channel subunit is expressed from a host cell.
- 30. 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.
- 31. The method of claim 30 wherein the labeled ligand for the α2δ-4 calcium channel subunit is a labeled gabapentin (GBP).
- 32. The method of claim 30 wherein said α2δ-4 calcium channel subunit is isolated or substantially purified.
- 33. The method of claim 30 wherein said α2δ-4 calcium channel subunit is associated with a membrane.
- 34. The method of claim 30 wherein said α2δ-4 calcium channel subunit is expressed from a host cell.
- 35. 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.
- 36. The method of claim 35 wherein the contacting step occurs in a cell.
- 37. The method of claim 35 wherein the reporter gene is the α2δ-4 gene.
BACKGROUND OF THE INVENTION
[0001] This application is a continuation-in-part of U.S. patent application Ser. No. 09/833,222 filed Apr. 11, 2001 and entitled “cDNA Encoding the Human Alpha2 Delta4 Calcium Channel Subunit”. The contents of which are hereby incorporated by reference in its entirety.
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
09833222 |
Apr 2001 |
US |
Child |
10119624 |
Apr 2002 |
US |