Interrogating the Druggability of the 2-Oxoglutarate-Dependent Dioxygenase Target Class by Chemical Proteomics

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• 作者：Gérard Joberty ; Markus Boesche ; Jack A. Brown ; Dirk Eberhard ; Neil S. Garton ; Philip G. Humphreys ; Toby Mathieson ; Marcel Muelbaier ; Nigel G. Ramsden ; Valérie Reader ; Anne Rueger ; Robert J. Sheppard ; Susan M. Westaway ; Marcus Bantscheff ; Kevin Lee ; David M. Wilson ; Rab K. Prinjha ; Gerard Drewes
• 刊名：ACS Chemical Biology
• 出版年：2016
• 出版时间：July 15, 2016
• 年：2016
• 卷：11
• 期：7
• 页码：2002-2010
• 全文大小：609K
• 年卷期：0
• ISSN：1554-8937

文摘

The 2-oxoglutarate-dependent dioxygenase target class comprises around 60 enzymes including several subfamilies with relevance to human disease, such as the prolyl hydroxylases and the Jumonji-type lysine demethylases. Current drug discovery approaches are largely based on small molecule inhibitors targeting the iron/2-oxoglutarate cofactor binding site. We have devised a chemoproteomics approach based on a combination of unselective active-site ligands tethered to beads, enabling affinity capturing of around 40 different dioxygenase enzymes from human cells. Mass-spectrometry-based quantification of bead-bound enzymes using a free-ligand competition-binding format enabled the comprehensive determination of affinities for the cosubstrate 2-oxoglutarate and for oncometabolites such as 2-hydroxyglutarate. We also profiled a set of representative drug-like inhibitor compounds. The results indicate that intracellular competition by endogenous cofactors and high active site similarity present substantial challenges for drug discovery for this target class.