Bacterial Intracellular Sodium Ion Measurement using CoroNa Green
Bacterial Intracellular Sodium Ion Measurement using CoroNa Green
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DOI：
10.21769/bioprotoc.2092
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发表时间：
2017-01-05
期刊：
影响因子：
0.8
通讯作者：
Minamino, Tohru
中科院分区：
文献类型：
--
作者：
Morimoto, Yusuke, V;Namba, Keiichi;Minamino, Tohru
细菌鞭毛 III 型输出装置由细胞质 ATP 酶复合物和跨膜输出门复合物组成，分别由穿过细胞质膜的 ATP 和质子动力 (PMF) 提供动力，并将鞭毛成分蛋白从细胞质转运到生长的鞭毛结构的远端，在那里进行组装 (Minamino, 2014)。当细胞质 ATP 酶复合物不能正常工作时，输出门复合物除了 PMF 之外还可以利用钠动力。跨膜输出门蛋白 FlhA 作为双离子通道传导 H+ 和 Na+ (Minamino et al., 2016)。在这里，我们描述了如何使用钠敏感荧光染料 CoroNa Green 测量活大肠杆菌细胞内的 Na+ 浓度（Minamino 等人，2016）。通过落射荧光显微镜测量 CoroNa Green 的荧光强度使我们能够定量测量细胞内 Na+ 浓度。
The bacterial flagellar type III export apparatus consists of a cytoplasmic ATPase complex and a transmembrane export gate complex, which are powered by ATP and proton motive force (PMF) across the cytoplasmic membrane, respectively, and transports flagellar component proteins from the cytoplasm to the distal end of the growing flagellar structure where their assembly occurs (Minamino, 2014). The export gate complex can utilize sodium motive force in addition to PMF when the cytoplasmic ATPase complex does not work properly. A transmembrane export gate protein FlhA acts as a dual ion channel to conduct both H+ and Na+ (Minamino et al., 2016). Here, we describe how to measure the intracellular Na+ concentrations in living Escherichia coli cells using a sodium-sensitive fluorescent dye, CoroNa Green (Minamino et al., 2016). Fluorescence intensity measurements of CoroNa Green by epi-fluorescence microscopy allows us to measure the intracellular Na+ concentration quantitatively.