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.
- 3. The isolated and purified DNA of claim 1, which encodes a human sphingosine kinase type 2 isoform.
- 4. An isolated and purified DNA selected from the group consisting of
- 5. An isolated and purified DNA selected from the group consisting of
- 6. A protein encoded by the DNA of claim 4.
- 7. An isolated protein comprising an amino acid sequence of murine SPHK2 consisting essentially of SEQ ID NO: 12.
- 8. An isolated and purified DNA which encodes a protein of a sphingosine kinase type 2 isoform, said DNA comprising a sequence selected from the group consisting of the sequence of Genbank Accession No. bankit325787 and the sequence of Genbank Accession No. bankit325752.
- 9. A recombinant DNA construct comprising:
(a) a vector and (b) the DNA of claim 2.
- 10. The recombinant DNA construct according to claim 9, wherein said vector is an expression vector.
- 11. The recombinant DNA construct according to claim 9, wherein said vector is a prokaryotic vector.
- 12. The recombinant DNA construct according to claim 9, wherein said vector is a eukaryotic vector.
- 13. A host cell transformed with the recombinant DNA construct according to claim 9.
- 14. The host cell according to claim 13, wherein said cell is prokaryotic.
- 15. The host cell according to claim 13, wherein said cell is eukaryotic.
- 16. A recombinant DNA construct comprising:
(a) a vector and (b) the DNA of claim 2.
- 17. The recombinant DNA construct according to claim 16, wherein said vector is an expression vector.
- 18. The recombinant DNA construct according to claim 16, wherein said vector is a prokaryotic vector.
- 19. The recombinant DNA construct according to claim 16, wherein said vector is a eukaryotic vector.
- 20. A host cell transformed with the recombinant DNA construct according to claim 16.
- 21. The host cell according to claim 20, wherein said cell is prokaryotic.
- 22. The host cell according to claim 20, wherein said cell is eukaryotic.
- 23. A method for producing a mouse sphingosine kinase type 2 isoform protein which comprises culturing the host cell according to claim 13, under conditions such that an isolated mouse sphingosine kinase type 2 isoform DNA is expressed and said mouse sphingosine kinase type 2 isoform protein is thereby produced.
- 24. A method for producing a human sphingosine kinase type 2 isoform protein which comprises culturing the host cell according to claim 20, under conditions such that an isolated human sphingosine kinase type 2 isoform DNA is expressed and said human sphingosine kinase type 2 isoform protein is thereby produced.
- 25. 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 9, 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.
- 26. An agent or drug detected by the method of claim 25.
- 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 16, 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. An agent or drug detected by the method of claim 27.
- 29. A method of regulating a biological process in a mammal comprising administering to a mammal in need thereof a pharmaceutically effective amount of the protein according to claim 6.
- 30. The method of claim 29, wherein the mammal is a human.
- 31. The method of claim 30, wherein the biological process is selected from the group consisting of mitogenesis, apoptosis, neuronal development, chemotaxis, angiogenesis and inflammatory responses.
- 32. The method of claim 30, wherein the biological process is angiogenesis.
- 33. 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 protein according to claim 6.
- 34. The method of claim 33, wherein the mammal is a human.
- 35. A method for the treatment or amelioration 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 protein according to claim 6.
- 36. The method of claim 35, wherein the mammal is a human.
- 37. 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 protein according to claim 6.
- 38. The method of claim 37, wherein the mammal is a human.
- 39. The method of claim 38, wherein the disease is cancer.
- 40. A composition for treating or ameliorating a disease resulting from increased cell death or decreased cell proliferation comprising a pharmaceutically effective amount of a protein according to claim 6, and a pharmaceutically acceptable carrier.
- 41. A method for the treatment or amelioration of developmental retardation in a human comprising administering to a human in need thereof a pharmaceutically effective amount of the protein according to claim 6.
- 42. 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.
- 43. An agent or a drug detected by the method according to claim 42.
- 44. 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.
- 45. A method for detecting sphingosine kinase type 2 in a sample comprising subjecting the sample to a polymerase chain reaction, a northern hybridization assay or an in situ hybridization assay and detecting for the presence of sphingosine kinase type 2.
- 46. A diagnostic kit for detecting sphingosine kinase type 2 RNA/cDNA in a sample comprising primers or nucleotides 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 RNA or cDNA and suitable ancillary reagents.
- 47. A method for detecting an agent or a drug which inhibits or promotes sphingosine kinase type 2 activity comprising:
(a) introducing a recombinant DNA construct comprising a vector and a DNA encoding a mammalian sphingosine kinase type 2 isoform in to 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 such 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.
- 48. The method for detecting an agent or a drug according to claim 47, wherein said mammalian sphingosine kinase type 2 isoform is mouse sphingosine kinase type 2 isoform.
- 49. The method for detecting an agent or a drug according to claim 47, wherein said mammalian sphingosine kinase type 2 isoform is mouse sphingosine kinase type 2 isoform.
- 50. An agent or a drug detected by the method according to claim 47.
- 51. An agent or a drug detected by the method according to claim 48.
- 52. An agent or a drug detected by the method according to claim 49.
- 53. A recombinant DNA construct comprising:
(a) a vector and (b) the DNA of claim 4.
- 54. A recombinant DNA construct comprising:
(a) a vector and (b) the DNA of claim 5.
- 55. An antibody specific for a protein according to claim 6.
- 56. The antibody according to claim 55, wherein the antibody is a polyclonal antibody.
- 57. The antibody according to claim 55, wherein the antibody is a monoclonal antibody.
- 58. A method of regulating a biological process in a mammal comprising administering to a mammal in need thereof a pharmaceutically effective amount of the protein according to claim 7.
- 59. The method of claim 57; wherein the mammal is a human.
- 60. 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 protein according to claim 7.
- 61. The method of claim 60, wherein the mammal is a human.
- 62. A method for the treatment or amelioration of a disease resulting from decreased cell death or increased cell proliferation comprising administering to a mammal in nee thereof a pharmaceutically effective amount of an antibody to a protein according to claim 7.
- 63. The method of claim 62, wherein the mammal is a human.
- 64. 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 protein according to claim 7.
- 65. The method of claim 64, wherein the mammal is a human.
- 66. An antibody specific for a protein according to claim 7.
- 67. The antibody according to claim 66, wherein the antibody is a polyclonal antibody.
- 68. The antibody according to claim 66, wherein the antibody is a monoclonal antibody.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Divisional application of application Ser. No. 09/817,676, filed Mar. 26, 2001, which claims the benefit under 35 USC 119(e) of Provisional Application Serial No. 60/194,318, filed Apr. 3, 2000.
GOVERNMENT RIGHTS
[0002] This invention was made with United States government support under Grant GM 43880 from the National Institutes of Health and a Postdoctoral Fellowship BC961968 from the United States Army Medical Research and Materiel Command, Prostate Cancer Research Program (VEN). The United States government has certain rights in this invention.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60194318 |
Apr 2000 |
US |
Divisions (1)
|
Number |
Date |
Country |
Parent |
09817676 |
Mar 2001 |
US |
Child |
10830677 |
Apr 2004 |
US |