PHARMACEUTICALS AND METHODS FOR TREATING HYPOXIA AND SCREENING METHODS THEREFOR

摘要

Light-generating fusion proteins having a ligand binding site and a light-generating polypeptide moiety and their use as diagnostics, in drug screening and discovery, and as therapeutics, are disclosed. The light-generating fusion protein has a feature where the bioluminescence of the polypeptide moiety changes upon binding of a ligand at the ligand binding site. The ligand may be, for example, an enzyme present in an environment only under certain conditions, e.g., ubiquitin ligase in a hypoxic state, such that the light-generating fusion protein is “turned on” only under such conditions.

主权项

1. A method of identifying prolyl hydroxylase modulators, comprising the steps of: a) contacting a prolyl hydroxylase, a putative prolyl hydroxylase modulator and a light-generating fusion protein comprising an HIFα polypeptide moiety having binding character for prolyl hydroxylase and a light-generating polypeptide moiety, wherein the light generation of said light-generating polypeptide moiety changes upon binding of a prolyl hydroxylase to said HIFα polypeptide moiety, under conditions favorable for binding of a prolyl hydroxylase to a HIFα to form a test sample; and b) determining the ability of said putative prolyl hydroxylase modulator to modulate prolyl hydroxylase by measuring the light generated in said test sample.