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  • 标题:An optical assay of the transport activity of ClC-7
  • 本地全文:下载
  • 作者:Ilaria Zanardi ; Giovanni Zifarelli ; Michael Pusch
  • 期刊名称:Scientific Reports
  • 电子版ISSN:2045-2322
  • 出版年度:2013
  • 卷号:3
  • DOI:10.1038/srep01231
  • 出版社:Springer Nature
  • 摘要:Osteoporosis, characterized by excessive osteoclast mediated bone resorption, affects millions of people worldwide representing a major public health problem. ClC-7 is a chloride-proton exchanger localized in lysosomes and in the resorption lacuna in osteoclasts where it is essential for bone resorption. Thus, drugs targeted at ClC-7 have been proposed for ameliorating osteoporosis. However, functional assays suited for high throughput screening (HTS) of ClC-7 function are lacking. Here we describe two complementary variants of purely optical assays of the transport activity of ClC-7, redirected to the plasma membrane employing a genetically encoded fluorescent Cl/pH indicator fused to the ClC-7 protein. These simple and robust functional assays of ClC-7 transport are well-suited to be applied in HTS of small-molecule inhibitors and may help to develop drugs suited for the treatment of osteoporosis.
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