Claims
- 1. An isolated and purified DNA which encodes a mammalian sphingosine kinase type 2 isoform.
- 2. The isolated and purified DNA of claim 1, which encodes a mouse sphingosine kinase type 2 isoform DNA.
- 3. The isolated and purified DNA of claim 1, which encodes a human sphingosine kinase type 2 isoform DNA.
- 4. The isolated and purified DNA of claim 2, which encodes a peptide of 617 amino acids.
- 5. The isolated and purified DNA of claim 3, which encodes a peptide of 618 amino acids.
- 6. A peptide encoded by the DNA of claim 4.
- 7. A peptide encoded by the DNA of claim 5.
- 8. An amino acid sequence of human SPHK2 consisting essentially of SEQ ID NO: 14.
- 9. An amino acid sequence of murine SPHK2 consisting essentially of SEQ ID NO: 12.
- 10. An isolated and purified DNA which encodes a peptide of a sphingosine kinase type 2 isoform, said DNA comprising a sequence selected from the group consisting of the sequence of Genbank Accession No. nkit325787 and the sequence of Genbank Accession No. bankit325752.
- 11. A recombinant DNA construct comprising:
(a) a vector and (b) the DNA of claim 2.
- 12. A recombinant DNA construct comprising:
(a) a vector and (b) the DNA of claim 3.
- 13. The recombinant DNA construct according to claim 11, wherein said vector is an expression vector.
- 14. The recombinant DNA construct according to claim 11, wherein said vector is a prokaryotic vector.
- 15. The recombinant DNA construct according to claim 11, wherein said vector is a eukaryotic vector.
- 16. The recombinant DNA construct according to claim 12, wherein said vector is an expression vector.
- 17. The recombinant DNA construct according to claim 12, wherein said vector is a prokaryotic vector.
- 18. The recombinant DNA construct according to claim 12, wherein said vector is a eukaryotic vector.
- 19. A host cell transformed with the recombinant DNA construct according to claim 11.
- 20. The host cell according to claim 19, wherein said cell is prokaryotic.
- 21. The host cell according to claim 19, wherein said cell is eukaryotic.
- 22. A host cell transformed with the recombinant DNA construct according to claim 12.
- 23. The host cell according to claim 22, wherein said cell is prokaryotic.
- 24. The host cell according to claim 22, wherein said cell is eukaryotic.
- 25. A method for producing a mouse sphingosine kinase type 2 isoform peptide which comprises culturing the host cell according to claim 19, under conditions such that an isolated mouse sphingosine kinase type 2 isoform DNA is expressed and said mouse sphingosine kinase type 2 isoform peptide is thereby produced.
- 26. A method for producing a human sphingosine kinase type 2 isoform peptide which comprises culturing the host cell according to claim 22, under conditions such that an isolated human sphingosine kinase type 2 isoform DNA is expressed and said human sphingosine kinase type 2 isoform peptide is thereby produced.
- 27. A method for detecting an agent or a drug which inhibits or promotes sphingosine kinase type 2 activity comprising:
(a) providing a recombinant DNA construct according to claim 11, into a cell such that sphingosine kinase type 2 isoform is produced in said cell; (b) adding at least one drug or agent to said cell, and (c) detecting whether or not said drug or agent inhibits or promotes sphingosine kinase type 2 activity by measuring sphingosine kinase-dependent phosphorylation of lipids in said cells and comparing the resultant measurement to a control which did not receive said drug or agent, wherein a decrease in the amount of sphingosine kinase-dependent phosphorylation of lipids as compared to the control indicates an inhibitory drug or agent, or an increase in the amount of sphingosine kinase-dependent phosphorylation of lipids in said cell as compared to the control indicates a stimulatory drug or agent.
- 28. A method for detecting an agent or a drug which inhibits or promotes sphingosine kinase type 2 activity comprising:
(a) providing a recombinant DNA construct according to claim 12, into a cell such that sphingosine kinase type 2 isoform is produced in said cell; (b) adding at least one drug or agent to said cell, and (c) detecting whether or not said drug or agent inhibits or promotes sphingosine kinase type 2 activity by measuring sphingosine kinase-dependent phosphorylation of lipids in said cells and comparing the resultant measurement to a control which did not receive said drug or agent, wherein a decrease in the amount of sphingosine kinase-dependent phosphorylation of lipids as compared to the control indicates an inhibitory drug or agent, or an increase in the amount of sphingosine kinase-dependent phosphorylation of lipids in said cell as compared to the control indicates a stimulatory drug or agent.
- 29. An agent or drug detected by the method of claim 27.
- 30. An agent or drug detected by the method of claim 28.
- 31. A method of regulating a biological process in a mammal comprising administering to a mammal in need thereof a pharmaceutically effective amount of the peptide according to claim 6.
- 32. A method of regulating a biological process in a mammal comprising administering to a mammal in need thereof a pharmaceutically effective amount of the peptide according to claim 7.
- 33. The method of claim 31, wherein the biological process is selected from the group consisting of mitogenesis, apoptosis, neuronal development, chemotaxis, angiogenesis and inflammatory responses.
- 34. The method of claim 31, wherein the biological process is angiogenesis.
- 35. The method of claim 32, wherein the biological process is selected from the group consisting of mitogenesis, apoptosis, neuronal development, chemotaxis, angiogenesis and inflammatory responses.
- 36. The method of claim 32, wherein the biological process is angiogenesis.
- 37. A method for the treatment or amelioration of a disease resulting from increased cell death or decreased cell proliferation, comprising administering to a mammal in need thereof a pharmaceutically effective amount of a peptide according to claim 6.
- 38. A method for the treatment or amelioration of a disease resulting from increased cell death or decreased cell proliferation, comprising administering to a mammal in need thereof a pharmaceutically effective amount of a peptide according to claim 7.
- 39. A method for the treatment or administration of a disease resulting from decreased cell death or increased cell proliferation comprising administering to a mammal in need thereof a pharmaceutically effective amount of an antibody to a peptide according to claim 6.
- 40. A method for the treatment or administration of a disease resulting from decreased cell death or increased cell proliferation comprising administering to a mammal in need thereof a pharmaceutically effective amount of an antibody to a peptide according to claim 7.
- 41. A method for the treatment or amelioration of a disease resulting from abnormal migration or motility of cells selected from the group consisting of cancer, restenosis and diabetic neuropathy, the method comprising administering to a mammal in need thereof, a pharmaceutically effective amount of an antibody to a peptide according to claim 6.
- 42. A method for the treatment or amelioration of a disease resulting from abnormal migration or motility of cells selected from the group consisting of cancer, restenosis and diabetic neuropathy, the method comprising administering to a mammal in need thereof, a pharmaceutically effective amount of an antibody to a peptide according to claim 7.
- 43. The method of claim 41, wherein the disease is cancer.
- 44. The method of claim 42, wherein the disease is cancer.
- 45. A composition for treating or ameliorating a disease resulting from increased cell death or decreased cell proliferation comprising a pharmaceutically effective amount of a peptide according to claim 6, and a pharmaceutically acceptable carrier.
- 46. A composition for treating or ameliorating a disease resulting from increased cell death or decreased cell proliferation comprising a pharmaceutically effective amount of a peptide according to claim 7, and a pharmaceutically acceptable carrier.
- 47. A method for screening agents or drugs which reduce or eliminate sphingosine kinase type 2 activity, the method comprising detecting a decrease in sphingosine kinase type 2 enzyme activity in the presence of said agent or drug.
- 48. A method for detecting the presence of a sphingosine kinase type 2 isoform in a sample comprising
(i) contacting a sample with antibodies which recognize sphingosine kinase type 2; and (ii) detecting the presence or absence of a complex formed between sphingosine kinase type 2 and antibodies specific therefor.
- 49. A method for detecting sphingosine kinase type 2 in a sample comprising subjecting the sample to a polymerase chain reaction and detecting for the presence of sphinosine kinase type 2.
- 50. A diagnostic kit for detecting sphingosine kinase type 2 RNA/cDNA in a sample comprising primers or oligonucleotides specific for sphingosine kinase type 2 RNA or cDNA suitable for hybridization to sphingosine kinase type 2 RNA or cDNA and/or amplification of sphingosine kinase type 2 sequences and suitable ancillary reagents.
CROSS-REFERENCE TO RELATED APPLICATION
[0001] The present application claims the benefit of Provisional Application Serial No. 60/194,318, filed Apr. 3, 2000, wherein priority under 35 USC 119(e) is claimed.
GOVERNMENT RIGHTS
[0002] This invention was made with U.S. government support under Grant GM 43880 from the national Institutes of health and a Postdoctoral Fellowship BC961968 from the U.S. Army Medical Research and Materiel Command, Prostate Cancer Research Program (VEN). The U.S. government has certain rights in this invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60194318 |
Apr 2000 |
US |