Combined proteomic and biochemical analyses redefine the consensus sequence requirement for epidermal growth factor-like domain hydroxylation

Natural and synthetic 2-oxoglutarate derivatives are substrates for oncogenic variants of human isocitrate dehydrogenase 1 and 2

“…Note that the chiral 2OG derivatives were racemates. We have previously shown that some of these 2OG derivatives are substrates and/or inhibitors of the human 2OG oxygenases aspartate/asparagine-β-hydroxylase (AspH) ( 47 , 48 , 49 ) and factor-inhibiting hypoxia inducible factor-α (FIH) ( 50 , 51 , 52 ), which catalyze the oxidations of specific residues in epidermal growth factor–like domains and ankyrin repeat domains/other proteins, respectively ( 22 , 23 ). Crystal structures of some of the 2OG derivatives, which were alternative substrates and/or inhibitors, in complex with AspH and FIH implies stereoselectivity in their binding modes ( 22 , 23 ).…”

5-Substituted Pyridine-2,4-dicarboxylate Derivatives Have Potential for Selective Inhibition of Human Jumonji-C Domain-Containing Protein 5

“…We have reported that some C3-substituted 2,4-PDCA derivatives efficiently inhibit the 2OG oxygenase aspartate/asparagine-β-hydroxylase (AspH), , which catalyzes the post-translational hydroxylation of specific aspartate and asparagine residues in epidermal growth factor-like domains (EGFDs), − and/or the JmjC histone N ε -dimethyllysine-specific demethylase 4E (KDM4E). ,, Both AspH and the KDM4 subfamily of JmjC KDMs are medicinal chemistry targets for the development of cancer therapeutics, and a KDM4 inhibitor is presently in clinical trials. ,− Considering that 2,4-PDCA ( 1 ) inhibits isolated recombinant JMJD5 ∼15-fold less efficiently than AspH and with similar efficiency to KDM4E (Table , entry i), it was important to improve the selectivity of 2,4-PDCA for JMJD5 inhibition in order to identify molecules for use in biological and functional assignment studies.…”

Structure-guided optimisation of N-hydroxythiazole-derived inhibitors of factor inhibiting hypoxia-inducible factor-α

“…To investigate the potential of BNS for selective FIH inhibition, its selectivity was investigated with respect to other 2OGdependent protein oxidising enzymes, that is PHD2, Jumonji-C domain-containing protein 5 (JMJD5), aspartate/asparagineb-hydroxylase (AspH) and KDM4A, using reported SPE-MS inhibition assays. 58,61,69,70 In addition to PHD2 and KDM4A, we included AspH and JMJD5 in our selectivity studies, because AspH, like FIH, catalyses protein aspartate-and asparagineresidue hydroxylation (although in epidermal growth factorlike domains), 71,72 and because JMJD5 is structurally closely related to FIH. 73,74 The results reveal that, in addition to being a potent PHD2 (IC 50 = 0.11 mM; Table 1, entry iii) and FIH inhibitor, BNS inhibits JMJD5 (IC 50 = 0.25 mM) and AspH (IC 50 = 3.36 mM), whereas inhibition of KDM4A was not observed (IC 50 > 100 mM).…”

“…High levels of selectivity were also achieved over JMJD5 (∼25-fold), KDM4A (>300-fold) and AspH (∼100-fold), the latter being of interest as AspH catalyses, like FIH, the b-hydroxylation of asparaginyl residues. 71,72 Importantly, MS studies with isolated recombinant human 2OG oxygenases reveal that the N-hydroxythiazole-based FIH inhibitors show a different selectivity prole for inhibiting 2OG oxygenases than does NOFD (Table 6), 77 an observation that reects the differences in their FIH binding modes, as observed in co-crystal structures with FIH (Fig. 2 and 4).…”

“…High levels of selectivity were also achieved over JMJD5 (∼25-fold), KDM4A (>300-fold) and AspH (∼100-fold), the latter being of interest as AspH catalyses, like FIH, the β-hydroxylation of asparaginyl residues. 71,72…”