Claims
- 1. An isolated nucleic acid molecule comprising a nucleotide sequence encoding a MEKK2 protein, said MEKK2 protein having an amino acid sequence at least 99.1% identical to the amino acid sequence of SEQ ID NO:2, wherein % identity is determined according to the LALIGN algorithm using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
- 2. The nucleic acid molecule of claim 1, which encodes a MEKK2 protein having an amino acid sequence at least 99.5% identical to the amino acid sequence of SEQ ID NO:2.
- 3. The nucleic acid molecule of claim 1, which encodes a MEKK2 protein having an amino acid sequence at least 99.9% identical to the amino acid sequence of SEQ ID NO:2.
- 4. An isolated nucleic acid molecule which encodes a MEKK2 protein comprising the amino acid sequence of SEQ ID NO:2.
- 5. An isolated nucleic acid molecule which comprises the nucleotide sequence of SEQ ID NO:3.
- 6. An isolated nucleic acid molecule encoding a MEKK2 protein comprising amino acids 356-616 of SEQ ID NO:2.
- 7. An isolated nucleic acid molecule encoding a MEKK2 fragment consisting essentially of amino acids 356-616 of SEQ ID NO:2.
- 8. An isolated nucleic acid molecule encoding a fusion protein comprising a MEKK2 fragment consisting essentially of amino acids 356-616 of SEQ ID NO:2 and at least one non-MEKK2 polypeptide.
- 9. A vector comprising the nucleic acid molecule of any one of claims 1-2 and 4-8.
- 10. The vector of claim 9, which is an expression vector.
- 11. A host cell containing the vector of claim 10.
- 12. A method for producing a MEKK2 protein comprising culturing the host cell of claim 11 in a suitable medium until a MEKK2 protein is produced.
- 13. The method of claim 12, further comprising isolating the MEKK2 protein from the medium or the host cell.
- 14. An isolated MEKK2 protein comprising an amino acid sequence at least 99.1% identical to the amino acid sequence of SEQ ID NO:2, wherein % identity is determined according to the LALIGN algorithm using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
- 15. An isolated protein comprising the amino acid sequence of SEQ ID NO:2.
- 16. The protein of claim 14, which comprises an amino acid sequence at least 99.5% identical to the amino acid sequence of SEQ ID NO:2.
- 17. The protein of claim 14, which comprises an amino acid sequence at least 99.9% identical to the amino acid sequence of SEQ ID NO:2.
- 18. An isolated protein encoded by a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO3.
- 19. An isolated protein comprising amino acids 356-616 of SEQ ID NO:2.
- 20. An isolated MEKK2 fragment consisting essentially of amino acids 356-616 of SEQ ID NO:2.
- 21. A fusion protein comprising a MEKK2 fragment consisting essentially of amino acids 356-616 of SEQ ID NO:2 and at least one non-MEKK2 polypeptide.
- 22. A fusion protein comprising the MEKK2 protein of any one of claims 14-15 and 17-18, operatively linked to a polypeptide other than MEKK2.
- 23. The MEKK2 protein of any one of claims 14-19, wherein said protein is phosphorylated.
- 24. The MEKK2 protein of claim 23, wherein said protein includes at least 1 mole of phosphate per mole of protein.
- 25. The MEKK2 protein of claim 23, wherein said protein includes at least 2 moles of phosphate per mole of protein.
- 26. An isolated antibody that specifically binds to the MEKK2 protein of claim 14.
- 27. An isolated antibody that specifically binds to the MEKK2 protein of claim 23.
- 28. The antibody of claim 26, which is a polyclonal antibody.
- 29. The antibodies of claim 26, which is a monoclonal antibody.
- 30. The antibody of claim 26, which is coupled to a detectable substance.
- 31. The antibody of claim 27, which is a polyclonal antibody.
- 32. The antibodies of claim 27, which is a monoclonal antibody.
- 33. The antibody of claim 27, which is coupled to a detectable substance.
- 34. A nonhuman transgenic animal that contains cells carrying a transgene that encodes the MEKK2 protein of claim 14.
- 35. A nonhuman transgenic animal that having the gene encoding the MEKK2 protein set forth as SEQ ID NO:2 mutated or deleted such that the animal fails to express the MEKK2 protein.
- 36. A method for specifically detecting the presence the MEKK2 protein of claim 11 in a biological sample comprising contacting the biological sample with an agent capable of specifically detecting the MEKK2 protein such that the presence of the MEKK2 protein is detected in the biological sample.
- 37. A method for specifically detecting the presence the nucleic acid molecule of claim 1 in a biological sample comprising contacting the biological sample with an agent capable of specifically detecting the nucleic acid molecule such that the presence of the nucleic acid molecule is detected in the biological sample.
- 38. A method for identifying a MEKK2 modulator comprising contacting the MEKK2 protein of claim 14 with a test compound; determining whether the compound modulates an activity of the MEKK2 protein; and identifying the compound as a MEKK2 modulator where the compound modulates the activity of the MEKK2 protein.
- 39. A method for identifying a MEKK2 modulator comprising contacting the MEKK2 protein of claim 14 with a test compound and determining the effect of the test compound on an activity of the MEKK2 protein to thereby identify the compound as a MEKK2 modulator.
- 40. A method for identifying a MEKK2 modulator comprising contacting a cell which expresses the MEKK2 protein of claim 14 with a test compound and determining the effect of the test compound on an activity of the MEKK2 protein to thereby identify the compound as a MEKK2 modulator.
- 41. The method of claim 39 or 40, wherein determining the effect of the test compound on an activity of the MEKK2 protein comprises determining a stimulatory effect.
- 42. The method of claim 39 or 40, wherein determining the effect of the test compound on an activity of the MEKK2 protein comprises determining an inhibitory effect.
- 43. The method of claim 40, wherein the cell further comprises a reporter gene responsive to the MEKK2 protein and determining whether the compound modulates an activity of the MEKK2 protein comprises determining the level of expression of the reporter gene in the presence of the test compound to thereby identify the MEKK2 modulator.
- 44. A method for identifying a MEKK2 modulator comprising contacting a cell which expresses the MEKK2 nucleic acid molecule of claim 1 with a test compound and determining the effect of the test compound on expression of the MEKK2 nucleic acid molecule to thereby identify the compound as a MEKK2 modulator.
- 45. The method of claim 44, wherein determining the effect of the test compound on expression of the MEKK2 nucleic acid molecule comprises determining a stimulatory effect.
- 46. The method of claim 44, wherein determining the effect of the test compound on expression of the MEKK2 nucleic acid molecule comprises determining an inhibitory effect.
- 47. A method for identifying a MEKK2-specific modulator comprising:
(a) contacting a MEKK2 protein or cell expressing said MEKK2 protein with a test compound; (b) determining the effect of the test compound on the expression or activity of the MEKK2; (c) contacting a MEKK1, MEKK3 or MEKK4 protein or cell expressing said MEKK1, MEKK3 or MEKK4 protein with said test compound; (d) determining the effect of the test compound on the expression or activity of the MEKK1, MEKK3 or MEKK4 protein; (e) comparing the effect as determined in subpart (b) with the effect as determined in subpart (d); and (f) identifying the compound as a MEKK2-specific modulator where said compound exhibits an effect as determined in subpart (b) and exhibits no effect as determined in subpart (d).
- 48. The method of any one of claims 38-40 and 44, wherein the test compound is selected from the group consisting of an antisense nucleic acid molecule, a peptide, a peptidomimetic or a small molecule.
- 49. The method of claim 39 or 40, wherein the activity is selected from the group consisting of (i) interaction of a MEKK2 protein with a MEKK2 binding partner, wherein the binding partner effects the activity of the MEKK2 molecule; (ii) interaction of a MEKK2 protein with a MEKK2 target molecule, wherein the MEKK2 protein effects the activity of the target molecule; (iii) phosphorylation of a MEKK2 target molecule selected from the group consisting of MKK1, MKK2, MKK3, MKK4, MKK5, MKK6, and MKK7; (iv) phosphorylation of a non-target protein; (v) mediation of activation of MAPK signal transduction molecules; (vi) modulation of the activity of a nuclear transcription factor; and (vii) modulation of ERK-, JNK- or p38-dependent gene transcription.
- 50. The method of claim 39 or 40, wherein the activity is selected from the group consisting of (i) modulation of cellular signal transduction, either in vitro or in vivo; (ii) regulation of gene transcription in a cell expressing a MEKK2 protein; (iii) regulation of gene transcription in a cell expressing a MEKK2 protein, wherein said cell is involved inflammation; (iv) regulation of cellular proliferation; (v) regulation of cellular differentiation; (vi) regulation of development; (vii) regulation of cell death; and (viii) regulation of inflammation.
- 51. Use of the MEKK2 protein set forth as SEQ ID NO:2 in a method for identifying a compound which specifically modulates the expression or activity of a non-MEKK2 protein, wherein a lack of expression or activity of said MEKK2 protein as compared to the expression or activity of the non-MEKK2 protein indicates that the compound specifically modulates the expression or activity of the non-MEKK2 protein.
- 52. A method for modulating a MEKK2 activity in a cell comprising contacting the cell with the MEKK2 modulator identified by the method of any one of claims 38-40 and 44, such that the MEKK2 activity in the cell is modulated.
RELATED APPLICATIONS
[0001] This application claims priority to U.S. provisional Application No. ______, formerly U.S. Ser. No. 10/163,811, filed on Jun. 5, 2002, the content of which is incorporated herein in its entirety by this reference.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60453712 |
Jun 2002 |
US |