A small-molecule probe for monitoring binding to prolyl hydroxylase domain 2 by fluorescence polarisation
Zhihong Li, Shuai Zhen, Kaijun Su, Anthony Tumber, Quanwei Yu, Ying Dong, M.A. McDonough, Christopher J. Schofield, Xiaojin Zhang
Abstract
Inhibition of the dioxygen sensing hypoxia-inducible factor prolyl hydroxylases has potential therapeutic benefit for treatment of diseases, including anaemia. We describe the discovery of a small-molecule probe useful for monitoring binding to human prolyl hydroxylase domain 2 (PHD2) via fluorescence polarisation. The assay is suitable for high-throughput screening of PHD inhibitors with both weak and strong affinities, as shown by work with clinically used inhibitors and naturally occurring PHD inhibitors.
Topics & Concepts
Small moleculeChemistryFluorescenceMoleculeCombinatorial chemistryBiophysicsBiochemistryBiologyOrganic chemistryPhysicsQuantum mechanicsUbiquitin and proteasome pathwaysCancer, Hypoxia, and MetabolismPeroxisome Proliferator-Activated Receptors