Claims
- 1. A method for screening for compounds that inhibit an activity of a target protein, wherein the target protein has microtubule stimulated ATPase activity and comprises a sequence that has greater than 90% amino acid identity to SEQ ID NO:2 or SEQ ID NO:4 as measured using a sequence comparison algorithm, the method comprising the steps of:contacting the target protein with a candidate agent at a first concentration and determining the target protein's ATPase activity; and contacting the target protein with a candidate agent at a second concentration and determining the target protein's ATPase activity; wherein a decrease in the level of ATPase activity of the target protein with an increase in concentration of the candidate agent indicates that the candidate agent inhibits the activity of the target protein.
- 2. A method of claim 1, wherein the screening occurs in a multi-well plate as part of a high-throughput screen.
- 3. A method of claim 1, wherein the target protein comprises an amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4.
- 4. A method of claim 1, wherein the target protein has greater than 95% amino acid sequence identity to SEQ ID NO:2 or SEQ ID NO:4 as measured using a sequence comparison algorithm.
- 5. A method of claim 1, wherein the target protein has greater than 98% amino acid sequence identity to SEQ ID NO:2 or SEQ ID NO:4 as measured using a sequence comparison algorithm.
- 6. A method of claim 1, wherein the target protein has been isolated from an endogenous source.
- 7. A method of claim 1, wherein the target protein has been produced recombinantly.
- 8. A method of claim 1, wherein the candidate agent is tested at zero concentration or at a level below detection.
- 9. A method of claim 1, wherein the candidate agent binds to the target protein.
- 10. A method of claim 1, wherein the target protein is contacted with the candidate agent in vitro.
- 11. A method of claim 1, wherein contacting the target protein with a candidate agent comprises adding the candidate agent to a mixture comprising the target protein under conditions which allow for the production of ADP or phosphate.
- 12. A method of claim 11, wherein determining the activity of the target protein comprises the steps of:i) subjecting the mixture to an enzymatic reaction, wherein said enzymatic reaction uses ADP or phosphate as a substrate under conditions which allow the ADP or phosphate to be utilized, and; ii) measuring NADH consumption as a measure of ADP production, wherein a decrease in NADH consumption with an increase in concentration of the candidate agent indicates that the candidate agent is an inhibitor of the target protein.
- 13. A method of claim 1, wherein the candidate agent is labeled.
- 14. A method for screening for compounds that activate an activity of a target protein, wherein the target protein has microtubule stimulated ATPase activity and comprises a sequence that has greater than 90% amino acid identity to SEQ ID NO:2 or SEQ ID NO:4 as measured using a sequence comparison algorithm, the method comprising the steps of:contacting the target protein with a candidate agent at a first concentration and determining the target protein's ATPase activity; and contacting the target protein with a candidate agent at a second concentration and determining the target protein's ATPase activity; wherein an increase in the level of ATPase activity of the target protein with an increase in concentration of the candidate agent indicates that the candidate agent activates the target protein.
- 15. A method of claim 14, wherein the screening occurs in a multi-well plate as part of a high-throughput screen.
- 16. A method of claim 14, wherein the target protein comprises an amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4.
- 17. A method of claim 14, wherein the target protein has greater than 95% amino acid sequence identity to SEQ ID NO:2 or SEQ ID NO:4 as measured using a sequence comparison algorithm.
- 18. A method of claim 14, wherein the target protein has greater than 98% amino acid sequence identity to SEQ ID NO:2 or SEQ ID NO:4 as measured using a sequence comparison algorithm.
- 19. A method of claim 14, wherein the target protein has been isolated from an endogenous source.
- 20. A method of claim 14, wherein the target protein has been produced recombinantly.
- 21. A method of claim 14, wherein the candidate agent is tested at zero concentration or at a level below detection.
- 22. A method of claim 14, wherein the candidate agent binds to the target protein.
- 23. A method of claim 14, wherein the target protein is contacted with the candidate agent in vitro.
- 24. A method of claim 14, wherein contacting the target protein with a candidate agent comprises adding the candidate agent to a mixture comprising the target protein under conditions which allow for the production of ADP or phosphate.
- 25. A method of claim 24, wherein determining the activity of the target protein comprises the steps of:i) subjecting the mixture to an enzymatic reaction, wherein said enzymatic reaction uses ADP or phosphate as a substrate under conditions which allow the ADP or phosphate to be utilized, and; ii) measuring NADH consumption as a measure of ADP production, wherein an increase in NADH consumption with an increase in concentration of the candidate agent indicates that the candidate agent is an activator of the target protein.
- 26. A method of claim 14, wherein the candidate agent is labeled.
CROSS-REFERENCE TO RELATED APPLICATION
This application is a continuation of U.S Application Ser. No 09/641,806, filed Aug. 17, 2000, U.S. Pat. No. 6,395,527 which is incorporated herein by reference.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
6248594 |
Tang et al. |
Jun 2001 |
B1 |
Non-Patent Literature Citations (3)
Entry |
Nakagawa, et al. (1997) Proc. Natl. Acad. Sci. U.S.A. 94 (18), 9654-9659 “Identification and classification of 16 new kinesin superfamily (KIF) proteins in mouse genome”. |
GenBank, Direct Submission, Accession No. AB001435, Mus musculus mRNA for motor domain of Kif7. |
GenPept, Direct Submission, Accession No. BAA22395, Motor domain of Kif7. |
Continuations (1)
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Number |
Date |
Country |
Parent |
09/641806 |
Aug 2000 |
US |
Child |
09/723129 |
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US |