Protease regulator screening assay
AssigneeChiron Corporation
InventorsDavid Y. Chien, Mark J. Selby, Kevin Shoemaker, Robert L. Warne
Life of this patent
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Abstract
A polypeptide containing an anchor region, a protease recognition site, and a detectable signal region can be produced recombinantly and directly attached to a solid support. The polypeptide is useful for screening protease regulators, especially protease inhibitors.
Claims3
A method for screening a compound capable of regulating protease activity comprising the steps of:
- incubating in the presence and absence of a test compound, a protease and a polypeptide wherein the polypeptide comprises an anchor region, a protease recognition site, and a detectable signal region, wherein the anchor region is bound to a solid support and the protease recognition site comprises a) a protease cleavable peptide bond, and b) at least 3 naturally adjacent amino acids on each side of the cleavable peptide bond,
- detecting the signal region bound to the solid support, wherein detecting an amount of the signal region bound to the solid support in the presence of the test compound which is greater or smaller than an amount detected in the absence of the test compound indicates that the test compound is capable of regulating protease activity.
A polypeptide consisting of a polyamino acid sequence, said polypeptide being useful for screening test compounds for protease regulating activity, said polypeptide comprising an anchor region, a protease recognition site, and a detectable signal region, wherein the anchor region is capable of binding to a solid support, and the protease recognition site comprises a) a cleavable peptide bond, and b) at least 3 naturally adjacent amino acids on each side of the cleavable peptide bond.
A method of forming a substrate for use in a screening assay for protease regulators comprising the steps of:
- binding a polypeptide consisting of a polyamino acid sequence to a solid support via a protein-protein interaction, wherein Kd of the protein-protein interaction is no more than 10−8, wherein the polypeptide comprises an anchor region, a protease recognition site, and a detectable signal region, wherein the protease recognition site comprises a) a cleavable peptide bond, and b) at least 3 naturally adjacent amino acids on each side of the cleavable peptide bond, and wherein the polypeptide binds to the solid support via the anchor region.
Dependencies are inferred from the claim text.