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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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