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Systematic Identification of MCU Modulators by Orthogonal Interspecies Chemical Screening
Molecular Cell ( IF 16.0 ) Pub Date : 2017-08-17 , DOI: 10.1016/j.molcel.2017.07.019
Daniela M. Arduino , Jennifer Wettmarshausen , Horia Vais , Paloma Navas-Navarro , Yiming Cheng , Anja Leimpek , Zhongming Ma , Alba Delrio-Lorenzo , Andrea Giordano , Cecilia Garcia-Perez , Guillaume Médard , Bernhard Kuster , Javier García-Sancho , Dejana Mokranjac , J. Kevin Foskett , M. Teresa Alonso , Fabiana Perocchi

The mitochondrial calcium uniporter complex is essential for calcium (Ca2+) uptake into mitochondria of all mammalian tissues, where it regulates bioenergetics, cell death, and Ca2+ signal transduction. Despite its involvement in several human diseases, we currently lack pharmacological agents for targeting uniporter activity. Here we introduce a high-throughput assay that selects for human MCU-specific small-molecule modulators in primary drug screens. Using isolated yeast mitochondria, reconstituted with human MCU, its essential regulator EMRE, and aequorin, and exploiting a D-lactate- and mannitol/sucrose-based bioenergetic shunt that greatly minimizes false-positive hits, we identify mitoxantrone out of more than 600 clinically approved drugs as a direct selective inhibitor of human MCU. We validate mitoxantrone in orthogonal mammalian cell-based assays, demonstrating that our screening approach is an effective and robust tool for MCU-specific drug discovery and, more generally, for the identification of compounds that target mitochondrial functions.



中文翻译:

正交物种间化学筛选系统识别MCU调制器。

线粒体钙单向转运体复合物对于所有哺乳动物组织的线粒体中的钙(Ca 2+)吸收都是必不可少的,在那里它调节生物能,细胞死亡和Ca 2+信号转导。尽管它参与了几种人类疾病,但我们目前缺乏靶向单向转运蛋白的药理剂。在这里,我们介绍了一种高通量检测方法,该方法可以在主要药物筛选中选择人类MCU特定的小分子调节剂。使用分离的酵母线粒体,并与人MCU,其必需的调节剂EMRE和水母发光蛋白重构,并利用基于D-乳酸和甘露醇/蔗糖的生物能分流器,可最大程度地降低假阳性率,我们从临床上鉴定出的米托蒽醌为600多种批准的药物可作为人类MCU的直接选择性抑制剂。我们在正交的基于哺乳动物细胞的实验中验证了米托蒽醌,这表明我们的筛选方法对于MCU特定药物的发现是一种有效而强大的工具,并且更广泛地说,

更新日期:2017-08-17
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