title: 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.
authors: Department of Chemistry and the Oxford Centre for Integrative Systems Biology, Chemistry Research Laboratory, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.|Pasteur Institute - Cenci Bolognetti Foundation, Department of Chemistry and Technologies of Drugs, University of Rome "La Sapienza", P.le A. Moro 5, 00185 Rome, Italy.
authors: Dante
authors: D
authors: Rotili
authors: Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
authors: Mikael
authors: M
authors: Altun
authors: Department of Chemistry and the Oxford Centre for Integrative Systems Biology, Chemistry Research Laboratory, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors: Akane
authors: A
authors: Kawamura
authors: Department of Chemistry and the Oxford Centre for Integrative Systems Biology, Chemistry Research Laboratory, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors: Alexander
authors: A
authors: Wolf
authors: Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
authors: Roman
authors: R
authors: Fischer
authors: Department of Chemistry and the Oxford Centre for Integrative Systems Biology, Chemistry Research Laboratory, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors: Ivanhoe K H
authors: IKH
authors: Leung
authors: Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
authors: Mukram M
authors: MM
authors: Mackeen
authors: Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
authors: Ya-Min
authors: YM
authors: Tian
authors: Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
authors: Peter J
authors: PJ
authors: Ratcliffe
authors: Pasteur Institute - Cenci Bolognetti Foundation, Department of Chemistry and Technologies of Drugs, University of Rome "La Sapienza", P.le A. Moro 5, 00185 Rome, Italy.
authors: Antonello
authors: A
authors: Mai
authors: Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
authors: Benedikt M
authors: BM
authors: Kessler
authors: Department of Chemistry and the Oxford Centre for Integrative Systems Biology, Chemistry Research Laboratory, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors: Christopher J
authors: CJ
authors: Schofield
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