Claims
- 1. A method of assaying for a candidate agent that modulates the attachment of a ubiquitin moiety to an Mdm2 protein, said method comprising:
a) combining:
i) a first ubiquitin agent comprising at least one ubiquitin moiety; ii) an Mdm2 protein; and iv) a candidate agent; and b) assaying for the attachment of said ubiquitin moiety to said Mdm2 protein.
- 2. The method according to claim 1, wherein said first ubiquitin agent is a ubiquitin conjugating agent.
- 3. The method according to claim 2, wherein step a) further comprises combining a ubiquitin activating agent comprising said ubiquitin moiety, thereby forming said ubiquitin conjugating agent comprising said ubiquitin moiety.
- 4. The method according to claim 2, wherein step a) further comprises combining a ubiquitin activating agent and said ubiquitin moiety, thereby forming said ubiquitin conjugating agent comprising said ubiquitin moiety.
- 5. A method of assaying for a candidate agent that modulates the attachment of a ubiquitin moiety to a p53 protein, said method comprising:
a) combining:
i) a ubiquitin conjugating agent comprising at least one ubiquitin moiety; ii) an mdm3 protein; iii) a p53 protein; and iv) a candidate agent; and b) assaying for the attachment of said ubiquitin moiety to said p53 protein.
- 6. The method according to claim 5, wherein step a) further comprises combining a ubiquitin conjugating agent and said ubiquitin moiety, thereby forming said ubiquitin conjugating agent comprising said ubiquitin moiety.
- 7. The method according to claim 5, wherein step a) further comprises combining a ubiquitin activating agent comprising said ubiquitin moiety, thereby forming said ubiquitin conjugating agent comprising said ubiquitin moiety.
- 8. The method according to claim 5, wherein step a) further comprises combining a ubiquitin activating agent and said ubiquitin moiety, thereby forming said ubiquitin conjugating agent comprising said ubiquitin moiety.
- 9. The method according to claim 1, wherein said Mdm2 protein comprises a first FRET label and said ubiquitin moiety comprises a second FRET label.
- 10. The method according to claim 1, wherein said Mdm2 protein comprises an attachment tag.
- 11. The method according to claim 10, wherein said Mdm2 protein is provided on a solid support.
- 12. The method according to claim 11, wherein said solid support comprises a microtiter plate.
- 13. The method according to claims 11, wherein said solid support comprises a bead.
- 14. The method according to claim 5, wherein said p53 protein comprises a first FRET label and said ubiquitin moiety comprises a second FRET label.
- 15. The method according to claim 5, wherein said p53 protein comprises an attachment tag.
- 16. The method according to claim 15, wherein said p53 protein is provided on a solid support.
- 17. The method according to claim 16, wherein said solid support comprises a microtiter plate.
- 18. The method according to claims 16, wherein said solid support comprises a bead.
- 19. The method according to any of claims 1-18, wherein said ubiquitin moiety comprises a label.
- 20. The method according to any of claims 1-18, wherein said ubiquitin moiety comprises an epitope label.
- 21. The method according to any of claims 1-18, wherein said combining comprises a first ubiquitin moiety comprising a first FRET label, and a second ubiquitin moiety comprising a second FRET label.
- 22. The method according to any of claims 1-18, wherein said Mdm2 protein is a human Mdm2 protein.
- 23. The method according to any of claims 1-18, wherein said ubiquitin moiety is a human ubiquitin.
- 24. The method according to any of claims 1-18, wherein said ubiquitin moiety is a ubiquitin derivative.
- 25. The method according to any of claims 1-18, wherein said candidate agent is a small molecule.
- 26. The method according to any of claims 1-18, wherein said candidate agent is a peptide.
- 27. A method of assaying for a candidate agent that modulates the attachment of a ubiquitin moiety to an Mdm2 protein, said method comprising:
a) combining:
i) a ubiquitin activating agent; ii) a ubiquitin conjugating agent; iii) an Mdm2 protein; iv) a candidate agent; and v) a ubiquitin moiety; and b) assaying for the attachment of said ubiquitin moiety to said Mdm2 protein.
- 28. A method of assaying for a candidate agent that modulates the attachment of a ubiquitin moiety to a p53 protein, said method comprising:
a) combining:
i) a ubiquitin activating agent; ii) a ubiquitin conjugating agent; iii) an Mdm2 protein; iv) a p53 protein v) a candidate agent; and vi) a ubiquitin moiety; and b) assaying for the attachment of said ubiquitin moiety to said p53 protein.
- 29. A method of assaying for a candidate agent that modulates the attachment of a second ubiquitin moiety to a p53 protein, said method comprising:
a) combining:
i) a ubiquitin activating agent; ii) a ubiquitin conjugating agent; iii) an Mdm2 protein; iv) a p53 protein comprising a first ubiquitin moiety, wherein said first ubiquitin moiety is labeled with a first FRET label; v) a candidate agent; and vi) a second ubiquitin moiety labeled with a second FRET label; and b) assaying for the attachment of said second ubiquitin moiety to said p53 protein by detecting a FRET reaction.
- 30. A method of assaying for a candidate agent that modulates the attachment of a first ubiquitin moiety to a p53 protein, said method comprising:
a) combining:
i) a ubiquitin conjugating agent comprising a first ubiquitin moiety labeled with a first FRET; ii) an Mdm2 protein; iii) a p53 protein comprising a second ubiquitin moiety, wherein said first ubiquitin moiety is labeled with a second FRET label; and iv) a candidate agent; and b) assaying for the attachment of said first ubiquitin moiety to said p53 protein by detecting a FRET reaction.
- 31. A method of assaying for a candidate agent that modulates the attachment of a first ubiquitin moiety to a p53 protein, said method comprising:
a) combining:
i) a ubiquitin activating agent comprising a first ubiquitin moiety labeled with a first FRET; ii) a ubiquitin conjugating agent; iii) an Mdm2 protein; iv) a p53 protein comprising a second ubiquitin moiety, wherein said first ubiquitin moiety is labeled with a second FRET label; and v) a candidate agent; and b) assaying for the attachment of said first ubiquitin moiety to said p53 protein by detecting a FRET reaction.
Parent Case Info
[0001] This application is a continuation-in-part application of U.S. patent application Ser. No. 09/826,312, filed Apr. 3, 2001 (pending) which is a continuation-in-part application U.S. patent application Ser. No. 09/542,487, filed Apr. 3, 2000 (pending).
Continuations (1)
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Number |
Date |
Country |
Parent |
10091139 |
Mar 2002 |
US |
Child |
10108767 |
Mar 2002 |
US |
Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
09826312 |
Apr 2001 |
US |
Child |
10091139 |
Mar 2002 |
US |
Parent |
09542487 |
Apr 2000 |
US |
Child |
09826312 |
Apr 2001 |
US |