Claims
- 1. A method for identifying a compound that modulates T lymphocyte activation, the method comprising the steps of:
(i) contacting the compound with a TRAC1 polypeptide or a fragment thereof, the polypeptide or fragment thereof encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide having an amino acid sequence of SEQ ID NO: 1; and (ii) determining the functional effect of the compound upon the TRAC1 polypeptide.
- 2. The method of claim 1, wherein the functional effect is measured in vitro.
- 3. The method of claim 2, wherein the functional effect is a physical effect.
- 4. The method of claim 2, wherein the functional effect is a chemical effect.
- 5. The method of claim 4, wherein the functional effect is determined by measuring ligase activity.
- 6. The method of claim 1, wherein the polypeptide is expressed in a host cell.
- 7. The method of claim 6, wherein the functional effect is a physical effect.
- 8. The method of claim 6, wherein the functional effect is a chemical or phenotypic effect.
- 9. The method of claim 6, wherein the host cell is primary T lymphocyte.
- 10. The method of claim 6, wherein the host cell is a cultured T cell.
- 11. The method of claim 10, wherein the host cell is a Jurkat cell.
- 12. The method of claim 6, wherein the chemical or phenotypic effect is determined by measuring CD69 expression, intracellular Ca2+ mobilization, Ca2+ influx, ligase activity, or lymphocyte proliferation.
- 13. The method of claim 1, wherein modulation is inhibition of T lymphocyte activation.
- 14. The method of claim 1, wherein the polypeptide is recombinant.
- 15. The method of claim 1, wherein the TRAC1 polypeptide comprises an amino acid sequence of SEQ ID NO: 1.
- 16. The method of claim 1, wherein the TRAC1 polypeptide is encoded by a nucleic acid comprising a nucleotide sequence of SEQ ID NO:2.
- 17. The method of claim 1, wherein the compound is an antibody.
- 18. The method of claim 1, wherein the compound is an antisense molecule.
- 19. The method of claim 1, wherein the compound is a small organic molecule.
- 20. The method of claim 1, wherein the compound is a peptide
- 21. The method of claim 20, wherein the peptide is circular.
- 22. A method for identifying a compound that modulates T lymphocyte activation, the method comprising the steps of:
(i) contacting a T cell comprising a TRAC1 polypeptide or fragment thereof with the compound, the TRAC1 polypeptide or fragment thereof encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide having an amino acid sequence of SEQ ID NO: 1; and (ii) determining the chemical or phenotypic effect of the compound upon the cell comprising the TRAC1 polypeptide or fragment thereof, thereby identifying a compound that modulates T lymphocyte activation.
- 23. A method for identifying a compound that modulates T lymphocyte activation, the method comprising the steps of:
(i) contacting the compound with a TRAC1 polypeptide or a fragment thereof, the TRAC1 polypeptide or fragment thereof encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide having an amino acid sequence of SEQ ID NO:1; (ii) determining the physical effect of the compound upon the TRAC1 polypeptide; and (iii) determining the chemical or phenotypic effect of the compound upon a cell comprising the TRAC1 polypeptide or fragment thereof, thereby identifying a compound that modulates T lymphocyte activation.
- 24. A method for identifying a compound capable of interfering with binding of an TRAC1 polypeptide or fragment thereof, the method comprising the steps of:
(i) combining an TRAC1 polypeptide or fragment thereof with an E2 ubiquitin-conjugating enzyme polypeptide and the compound, wherein the TRAC1 polypeptide or fragment thereof is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide having an amino acid sequence of SEQ ID NO:2; and (ii) determining the binding of the TRAC1 polypeptide or fragment thereof to the E2 ubiquitin-conjugating enzyme polypeptide.
- 25. The method of claim 24, wherein the TRAC1 polypeptide or fragment thereof has ligase activity.
- 26. The method of claim 24, wherein the E2 ubiquitin-conjugating enzyme polypeptide is selected from the group consisting of Ubc5, Ubc7, and Ubc8.
- 27. The method of claim 24, wherein the TRAC1 polypeptide or fragment thereof and the E2 ubiquitin-conjugating enzyme polypeptide are combined first.
- 28. The method of claim 24, wherein the reaction is performed in vitro.
- 29. The method of claim 24, wherein the TRAC1 polypeptide or fragment thereof and the E2 ubiquitin-conjugating enzyme polypeptide are expressed in a cell.
- 30. The method of claim 29, wherein the cell is a yeast cell.
- 31. The method of claim 30, wherein the TRAC1 polypeptide or fragment thereof is fused to a heterologous polypeptide.
- 32. The method of claim 24, wherein the binding of the TRAC1 polypeptide or fragment thereof to the E2 ubiquitin-conjugating enzyme polypeptide is determined by measuring reporter gene expression.
- 33. An isolated complex comprising a TRAC1 polypeptide or fragment thereof bound to an E2 ubiquitin-conjugating enzyme polypeptide, wherein the TRAC1 polypeptide or fragment thereof is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide having an amino acid sequence of SEQ ID NO:2.
- 34. The complex of claim 33, wherein the E2 ubiquitin-conjugating enzyme polypeptide is selected from the group consisting of Ubc5, Ubc7, and Ubc8.
- 35. A method of modulating T lymphocyte activation in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of a compound identified using the method of claim 1.
- 36. The method of claim 35, wherein the subject is a human.
- 37. The method of claim 35, wherein the compound is an antibody.
- 38. The method of claim 35, wherein the compound is an antisense molecule.
- 39. The method of claim 35, wherein the compound is a small organic molecule.
- 40. The method of claim 35, wherein the compound is a peptide.
- 41. The method of claim 40, wherein the peptide is circular.
- 42. The method of claim 35, wherein the compound inhibits T lymphocyte activation.
- 43. A method of modulating T lymphocyte activation in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of a TRAC1 polypeptide, the polypeptide encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide having an amino acid sequence of SEQ ID NO:1.
- 44. The method of claim 43, wherein the TRAC1 polypeptide comprises an amino acid sequence of SEQ ID NO:1.
- 45. A method of modulating T lymphocyte activation in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of a nucleic acid encoding a TRAC1 polypeptide, wherein the nucleic acid hybridizes under stringent conditions to a nucleic acid encoding a polypeptide having an amino acid sequence of SEQ ID NO:1.
- 46. The method of claim 45, wherein the TRAC1 nucleic acid comprises a nucleotide sequence of SEQ ID NO:2.
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] The present application claims priority to U.S. Ser. No. 60/282,432, filed Apr. 6, 2001, herein incorporated by reference in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60282432 |
Apr 2001 |
US |