Identifying molecules modulating protein-protein interactions using protease activated reporters

摘要

Assay methods and systems use enzymatic cleavage resulting from protein-protein interaction to modulate (activate or inactivate) a reporter.

主权项

1. An assay system to identify a compound that modulates a protein-protein interaction between a first protein and a second protein comprising:
i) a first protein that is attached to a reporter activating protein that is arranged to be inactive, wherein the inactive reporter activating protein comprises a cleavage site for a protease that is interposed between two portions of the inactive reporter activating protein; ii) a second protein that is attached to a protease, wherein the protease is capable of cleaving the cleavage site in the inactive reporter activating protein; and iii) a reporter whose signal is changed by the activity of an active reporter activating protein; wherein association of the first protein with the second protein causes the protease to cleave the inactive reporter activating protein at the cleavage site such that the two portions of the inactive reporter activating protein are rearranged to be active and thereby generate the active reporter activating protein.