Zhang Yiming, Zheng Sisi, Zhou Yubin, Gill Donald L, Wang Youjun
Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, China.
Department of Medical Physiology, Institute of Biosciences and Technology, Texas A&M University, Houston, TX, USA.
Methods Mol Biol. 2018;1843:1-16. doi: 10.1007/978-1-4939-8704-7_1.
Induced by the depletion of ER calcium store, the calcium influx through calcium release-activated calcium (CRAC) channels is an ubiquitous way of Ca influx for most cell types. This process is mediated by STIM protein, ER calcium sensor, and PM localized Orai calcium channels. Biophysical characterization of this STIM-Orai-mediated current, or I, with whole-cell patch-clamp technique is essential for revealing the molecular mechanisms underlying the process of STIM-Orai activation or modulation. Here we describe one commonly used procedure of monitoring CRAC activity with whole-cell patch-clamp technique.
内质网钙库耗竭所诱导的、通过钙释放激活钙（CRAC）通道的钙内流，是大多数细胞类型中普遍存在的钙内流方式。这一过程由内质网钙传感器STIM蛋白和定位在质膜的Orai钙通道介导。采用全细胞膜片钳技术对这种由STIM-Orai介导的电流（即I ）进行生物物理特性分析，对于揭示STIM-Orai激活或调节过程的分子机制至关重要。在此，我们描述一种使用全细胞膜片钳技术监测CRAC活性的常用方法。
