Bacterial Expression and HTS Assessment of Soluble Epoxide Hydrolase Phosphatase
Bacterial Expression and HTS Assessment of Soluble Epoxide Hydrolase Phosphatase
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DOI：
10.1177/1087057116637609
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发表时间：
2016-08-01
影响因子：
--
通讯作者：
Proschak, Ewgenij
中科院分区：
文献类型：
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作者：
Klingler, Franca-Maria;Wolf, Markus;Proschak, Ewgenij
可溶性环氧化物水解酶（sEH）是一种双功能酶，具有环氧化物水解酶和脂质磷酸酶活性（sEH-P）。虽然环氧化物水解酶结构域的生理作用是很好的理解，磷酸酶活性的后果仍然不清楚。在这里，我们描述了细菌表达的重组N-末端结构域的sEH-P和开发的高通量筛选方案，使用敏感的和商业上可获得的底物荧光素二磷酸。证明了测定系统的可用性，并鉴定了sEH-P的新型抑制剂。
Soluble epoxide hydrolase (sEH) is a bifunctional enzyme that possesses an epoxide hydrolase and lipid phosphatase activity (sEH-P) at two distinct catalytic domains. While the physiological role of the epoxide hydrolase domain is well understood, the consequences of the phosphatase activity remain unclear. Herein we describe the bacterial expression of the recombinant N-terminal domain of sEH-P and the development of a high-throughput screening protocol using a sensitive and commercially available substrate fluorescein diphosphate. The usability of the assay system was demonstrated and novel inhibitors of sEH-P were identified.