ASSAY FOR INHIBITORS OF CIP/KIP PROTEIN DEGRADATION

摘要

An assay and system compatible with high throughput screening (HTS) that is capable of identifying inhibitors, such as small-molecule inhibitors, of the degradation of the Cdk inhibitor p21, are described. The assay is based on the use of fusion protein comprising (i) a p2 polypeptide; and (i) a reporter protein linked to the C-terminal of said p21 polypeptide, wherein the fusion protein has a half-life that is similar to that of the p21 polypeptide. Inhibitors identified by this assay may be useful to inhibit the proliferation of tumor cells, and thus for the treatment of cancers.

主权项

1. A high throughput Screening (HTS)-compatible method for determining whether a test compound may be useful for treating cancer, said method comprising
(a) contacting said test compound with a cell expressing a fusion protein in the presence of a protein synthesis inhibitor, said fusion protein comprising (i) a Cip/Kip polypeptide; and (i) a reporter protein linked to the C-terminal of said Cip/Kip polypeptide, wherein said fusion protein has a half-life that is similar to that of said Cip/Kip polypeptide, and (b) measuring a readout signal from the reporter protein,wherein a higher readout signal from the reporter protein in the presence of said test compound, relative to the readout signal in the absence of said test compound, is indicative that said test compound may be useful for treating cancer.