Maitra Rangan, Sivashanmugam Perumal, Warner Keith
1RTI International, Research Triangle Park, NC, USA.
J Biomol Screen. 2013 Oct;18(9):1132-7. doi: 10.1177/1087057113488420. Epub 2013 May 7.
The cystic fibrosis transmembrane conductance regulator (CFTR) protein is an important regulator of ion transport and fluid secretion in humans. Mutations to CFTR cause cystic fibrosis, which is a common recessive genetic disorder in Caucasians. Involvement of CFTR has been noted in other important diseases, such as secretory diarrhea and polycystic kidney disease. The assays to monitor CFTR function that have been described to date either are complicated or require specialized instrumentation and training for execution. In this report, we describe a rapid FlexStation-based membrane potential assay to monitor CFTR function. In this assay, agonist-mediated activation of CFTR results in membrane depolarization that can be monitored using a fluorescent membrane potential probe. Availability of a simple mix-and-read assay to monitor the function of this important protein might accelerate the discovery of CFTR ligands to study a variety of conditions.
囊性纤维化跨膜传导调节因子（CFTR）蛋白是人体离子转运和液体分泌的重要调节因子。CFTR的突变会导致囊性纤维化，这是白种人中常见的隐性遗传病。CFTR还与其他重要疾病有关，如分泌性腹泻和多囊肾病。迄今为止所描述的用于监测CFTR功能的检测方法要么复杂，要么需要专门的仪器设备和操作培训。在本报告中，我们描述了一种基于FlexStation的快速膜电位检测方法来监测CFTR功能。在该检测中，激动剂介导的CFTR激活会导致膜去极化，可使用荧光膜电位探针进行监测。一种用于监测这种重要蛋白质功能的简单混合读取检测方法的出现，可能会加速CFTR配体的发现，以研究各种病症。
