Compositions and assays utilizing ADP or phosphate for detecting protein modulators
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Abstract
Described herein are methods which identify candidate agents as binding to a protein or as a modulator of the binding characteristics or biological activity of a protein. Generally, the methods involve the use of ADP or phosphate. The assays can be used in a high throughput system to obviate the cumbersome steps of using gels or radioactive materials.
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Citations
39 Claims
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1-29. -29. (canceled)
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30. A method of modulating polymerization or depolymerization of a cytoskeletal filament protein, wherein the filament protein comprises at least one of tubulin and actin, the method comprising providing a compound that modulates polymerization or depolymerization of the cytoskeletal filament protein, wherein the compound is characterized by
a) adding a test compound to a mixture comprising the filament protein under conditions which normally promote either polymerization or depolymerization of the filament protein; -
b) adding a motor protein to the mixture, wherein the motor protein has ATPase activity, and the ATPase activity is activated by polymerized filament protein, and wherein the motor protein is added to the mixture under conditions which normally allow the production of ADP or phosphate by said motor protein; and
c) determining the ATPase activity of the motor protein by detecting the formation of ADP or phosphate, wherein a change in ATPase activity in the presence of the compound compared to the absence of said compound is an indication that the test compound is suitable for use as the modulator of filament protein polymerization or depolymerization. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39)
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Specification