一种用于2-氧代戊二酸加氧酶的光反应性小分子探针。
A photoreactive small-molecule probe for 2-oxoglutarate oxygenases.
作者信息
Rotili Dante, Altun Mikael, Kawamura Akane, Wolf Alexander, Fischer Roman, Leung Ivanhoe K H, Mackeen Mukram M, Tian Ya-Min, Ratcliffe Peter J, Mai Antonello, Kessler Benedikt M, Schofield Christopher J
机构信息
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.
出版信息
Chem Biol. 2011 May 27;18(5):642-654. doi: 10.1016/j.chembiol.2011.03.007.
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.
2-氧代戊二酸（2-OG）依赖性加氧酶在人类生物学中具有多种作用。几种2-OG加氧酶的抑制作用正被作为治疗干预的靶点，包括用于癌症、贫血和缺血性疾病。我们报道了一种用于2-OG加氧酶的小分子探针，该探针采用与光可活化交联基团和亲和纯化标签偶联的羟基喹啉模板。在用重组蛋白进行研究后，该探针被证明可与人类粗细胞提取物中的2-OG加氧酶交联，包括与内源性水平的蛋白质交联。这种方法对于抑制剂分析很有用，如在HEK293T核提取物中与组蛋白去甲基化酶FBXL11（KDM2A）交联所证明的那样。结果还表明小分子探针可能适用于底物鉴定研究。
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