Reporter Ligand NMR Screening Method for 2‑Oxoglutarate Oxygenase Inhibitors

• Published in: Journal of medicinal chemistry Vol. 56; no. 2; pp. 547 - 555
• Main Authors: Leung, Ivanhoe K. H, Demetriades, Marina, Hardy, Adam P, Lejeune, Clarisse, Smart, Tristan J, Szöllössi, Andrea, Kawamura, Akane, Schofield, Christopher J, Claridge, Timothy D. W
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 24.01.2013
• Subjects: Citric Acid Cycle; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Humans; Ketoglutaric Acids - metabolism; Ligands; Magnetic Resonance Spectroscopy - methods; Oxygenases - antagonists & inhibitors
• ISSN: 0022-2623; 1520-4804
• DOI: 10.1021/jm301583m

The human 2-oxoglutarate (2OG) dependent oxygenases belong to a family of structurally related enzymes that play important roles in many biological processes. We report that competition-based NMR methods, using 2OG as a reporter ligand, can be used for quantitative and site-specific screening of ligand binding to 2OG oxygenases. The method was demonstrated using hypoxia inducible factor hydroxylases and histone demethylases, and K D values were determined for inhibitors that compete with 2OG at the metal center. This technique is also useful as a screening or validation tool for inhibitor discovery, as exemplified by work with protein-directed dynamic combinatorial chemistry.