钙传感器G-CatchER介导的快速亚细胞钙反应及动力学
Rapid subcellular calcium responses and dynamics by calcium sensor G-CatchER.
作者信息
Reddish Florence N, Miller Cassandra L, Deng Xiaonan, Dong Bin, Patel Atit A, Ghane Mohammad A, Mosca Barbara, McBean Cheyenne, Wu Shengnan, Solntsev Kyril M, Zhuo You, Gadda Giovanni, Fang Ning, Cox Daniel N, Mabb Angela M, Treves Susan, Zorzato Francesco, Yang Jenny J
机构信息
Department of Chemistry, Center for Diagnostics and Therapeutics, Advanced Translational Imaging Facility, Georgia State University, Atlanta, GA 30303, USA.
Neuroscience Institute, Georgia State University, Atlanta, GA 30303, USA.
出版信息
iScience. 2021 Feb 3;24(3):102129. doi: 10.1016/j.isci.2021.102129. eCollection 2021 Mar 19.
The precise spatiotemporal characteristics of subcellular calcium (Ca) transients are critical for the physiological processes. Here we report a green Ca sensor called "G-CatchER" using a protein design to report rapid local ER Ca dynamics with significantly improved folding properties. G-CatchER exhibits a superior Ca on rate to G-CEPIA1er and has a Ca-induced fluorescence lifetimes increase. G-CatchER also reports agonist/antagonist triggered Ca dynamics in several cell types including primary neurons that are orchestrated by IPRs, RyRs, and SERCAs with an ability to differentiate expression. Upon localization to the lumen of the RyR channel (G-CatchER-JP45), we report a rapid local Ca release that is likely due to calsequestrin. Transgenic expression of G-CatchER in muscle demonstrates its utility as an reporter of stimulus-evoked SR local Ca dynamics. G-CatchER will be an invaluable tool to examine local ER/SR Ca dynamics and facilitate drug development associated with ER dysfunction.
亚细胞钙（Ca）瞬变的精确时空特征对于生理过程至关重要。在此，我们报告一种名为“G-CatchER”的绿色钙传感器，它采用蛋白质设计来报告快速的局部内质网钙动态，其折叠特性有显著改善。G-CatchER对钙的结合速率优于G-CEPIA1er，并且钙诱导荧光寿命增加。G-CatchER还能报告激动剂/拮抗剂触发的几种细胞类型中的钙动态，包括由肌醇1,4,5-三磷酸受体（IPRs）、兰尼碱受体（RyRs）和肌浆网钙ATP酶（SERCAs）协调的原代神经元中的钙动态，且具有区分表达的能力。当定位于兰尼碱受体通道腔（G-CatchER-JP45）时，我们报告了一种可能由钙网蛋白引起的快速局部钙释放。G-CatchER在肌肉中的转基因表达证明了其作为刺激诱发的肌浆网局部钙动态报告分子的效用。G-CatchER将成为研究局部内质网/肌浆网钙动态以及促进与内质网功能障碍相关药物开发的宝贵工具。
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