title: Interrogating the Druggability of the 2-Oxoglutarate-Dependent Dioxygenase Target Class by Chemical Proteomics.
abstract: 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.
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Gérard
authors.initials: G
authors.lastname: Joberty
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Boesche
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: Jack A
authors.initials: JA
authors.lastname: Brown
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Eberhard
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: Neil S
authors.initials: NS
authors.lastname: Garton
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: Philip G
authors.initials: PG
authors.lastname: Humphreys
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Toby
authors.initials: T
authors.lastname: Mathieson
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Marcel
authors.initials: M
authors.lastname: Muelbaier
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Nigel G
authors.initials: NG
authors.lastname: Ramsden
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Valérie
authors.initials: V
authors.lastname: Reader
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Rueger
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Sheppard
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: Susan M
authors.initials: SM
authors.lastname: Westaway
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Bantscheff
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: Kevin
authors.initials: K
authors.lastname: Lee
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: David M
authors.initials: DM
authors.lastname: Wilson
authors.affiliation: Epinova Discovery Performance Unit, Medicines Research Centre, GlaxoSmithKline R&D , Stevenage, United Kingdom.
authors.forename: Rab K
authors.initials: RK
authors.lastname: Prinjha
authors.affiliation: Cellzome GmbH, a GlaxoSmithKline company , Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Gerard
authors.initials: G
authors.lastname: Drewes
mesh_terms: D049308:Dioxygenases / Q000378:metabolism*; D007656:Ketoglutaric Acids / Q000378:metabolism*; D040901:Proteomics
chemical_list: D007656:Ketoglutaric Acids; D049308:Dioxygenases