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A photoreactive small-molecule probe for 2-oxoglutarate oxygenases.

ABSTRACT: 2-oxoglutarate (2-OG)-dependent oxygenases have diverse roles in human biology. The inhibition of several 2-OG oxygenases is being targeted for therapeutic intervention, including for cancer, anemia, and ischemic diseases. We report a small-molecule probe for 2-OG oxygenases that employs a hydroxyquinoline template coupled to a photoactivable crosslinking group and an affinity-purification tag. Following studies with recombinant proteins, the probe was shown to crosslink to 2-OG oxygenases in human crude cell extracts, including to proteins at endogenous levels. This approach is useful for inhibitor profiling, as demonstrated by crosslinking to the histone demethylase FBXL11 (KDM2A) in HEK293T nuclear extracts. The results also suggest that small-molecule probes may be suitable for substrate identification studies.

SUBMITTER: Rotili D

PROVIDER: S-EPMC4681097 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature

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A photoreactive small-molecule probe for 2-oxoglutarate oxygenases.

Rotili Dante D Altun Mikael M Kawamura Akane A Wolf Alexander A Fischer Roman R Leung Ivanhoe K H IKH Mackeen Mukram M MM Tian Ya-Min YM Ratcliffe Peter J PJ Mai Antonello A Kessler Benedikt M BM Schofield Christopher J CJ

Chemistry & biology 20110501 5

2-oxoglutarate (2-OG)-dependent oxygenases have diverse roles in human biology. The inhibition of several 2-OG oxygenases is being targeted for therapeutic intervention, including for cancer, anemia, and ischemic diseases. We report a small-molecule probe for 2-OG oxygenases that employs a hydroxyquinoline template coupled to a photoactivable crosslinking group and an affinity-purification tag. Following studies with recombinant proteins, the probe was shown to crosslink to 2-OG oxygenases in hu ...[more]

PMID: 21609845

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Protein Hydroxylation Catalyzed by 2-Oxoglutarate-dependent Oxygenases.

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Catalytic Mechanisms of Fe(II)- and 2-Oxoglutarate-dependent Oxygenases.

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Amazing Diversity in Biochemical Roles of Fe(II)/2-Oxoglutarate Oxygenases.

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2-Oxoglutarate derivatives can selectively enhance or inhibit the activity of human oxygenases.

Project description:2-Oxoglutarate (2OG) oxygenases are validated agrochemical and human drug targets. The potential for modulating their activity with 2OG derivatives has not been explored, possibly due to concerns regarding selectivity. We report proof-of-principle studies demonstrating selective enhancement or inhibition of 2OG oxygenase activity by 2-oxo acids. The human 2OG oxygenases studied, factor inhibiting hypoxia-inducible transcription factor HIF-α (FIH) and aspartate/asparagine-β-hydroxylase (AspH), catalyze C3 hydroxylations of Asp/Asn-residues. Of 35 tested 2OG derivatives, 10 enhance and 17 inhibit FIH activity. Comparison with results for AspH reveals that 2OG derivatives selectively enhance or inhibit FIH or AspH. Comparison of FIH structures complexed with 2OG derivatives to those for AspH provides insight into the basis of the observed selectivity. 2-Oxo acid derivatives have potential as drugs, for use in biomimetic catalysis, and in functional studies. The results suggest that the in vivo activity of 2OG oxygenases may be regulated by natural 2-oxo acids other than 2OG.

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The function and catalysis of 2-oxoglutarate-dependent oxygenases involved in plant flavonoid biosynthesis.

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Ultrasensitive small molecule fluorogenic probe for human heparanase.

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Pathway from N-Alkylglycine to Alkylisonitrile Catalyzed by Iron(II) and 2-Oxoglutarate-Dependent Oxygenases.

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