title: Affinity-Based Fluorescence Polarization Assay for High-Throughput Screening of Prolyl Hydroxylase 2 Inhibitors.
abstract: Prolyl hydroxylase domain 2 (PHD2) enzyme, a Fe(II) and 2-oxoglutarate (2-OG) dependent oxygenase, mediates key physiological responses to hypoxia by modulating the levels of hypoxia inducible factor 1-α (HIF1α). PHD2 has been shown to have the therapeutic potentials for conditions including anemia and ischemic disease. Currently, many activity-based assays have been developed for identifying PHD2 inhibitors. Here we report an affinity-based fluorescence polarization method using FITC-labeled HIF1α (556-574) peptide as a probe for quantitative and site-specific screening of small molecule PHD2 inhibitors.
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Lei
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Tianhan
authors.initials: T
authors.lastname: Hu
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Xingsen
authors.initials: X
authors.lastname: Wu
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Qichao
authors.initials: Q
authors.lastname: Bao
authors.affiliation: National Engineering and Research Center for Target Drugs , Lianyungang 222000, China.
authors.forename: Lianshan
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Xia
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Haopeng
authors.initials: H
authors.lastname: Sun
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Qidong
authors.initials: Q
authors.lastname: You
authors.affiliation: Jiangsu Key Laboratory of Drug Design and Optimization, and State Key Laboratory of Natural Medicines, China Pharmaceutical University , Nanjing 210009, China ; Department of Organic Chemistry, School of Science, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Xiaojin
authors.initials: X
authors.lastname: Zhang
keywords: FITC-labeled probe; PHD2; affinity-based; fluorescence polarization assay