A genetically encoded toolkit for tracking live-cell histidine dynamics in space and time.
A genetically encoded toolkit for tracking live-cell histidine dynamics in space and time.
复制标题
用于跟踪活细胞组氨酸在空间和时间上的动态的基因编码工具包
DOI：
10.1038/srep43479
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发表时间：
2017-03-02
影响因子：
4.6
通讯作者：
Yang Y
中科院分区：
文献类型：
--
作者：
Hu H;Gu Y;Xu L;Zou Y;Wang A;Tao R;Chen X;Zhao Y;Yang Y
高分辨率的组氨酸在单个活的哺乳动物细胞的时空成像面临着技术挑战。在这里，我们开发了一系列的比率，高响应，单荧光蛋白为基础的组氨酸传感器的宽动态范围。我们使用这些传感器来量化葡萄糖剥夺细胞和葡萄糖喂养细胞中的亚细胞游离组氨酸浓度。结果表明，胞浆游离组氨酸浓度比线粒体中的游离组氨酸浓度高，对环境的反应更敏感。此外，组氨酸很容易跨质膜和线粒体内膜，这几乎有相似的运输速率和运输常数，组氨酸的运输不受细胞代谢状态的影响。这些传感器是跟踪亚细胞器内组氨酸动力学的潜在工具，它们将为探索复杂的组氨酸信号转导开辟一条途径。
High-resolution spatiotemporal imaging of histidine in single living mammalian cells faces technical challenges. Here, we developed a series of ratiometric, highly responsive, and single fluorescent protein-based histidine sensors of wide dynamic range. We used these sensors to quantify subcellular free-histidine concentrations in glucose-deprived cells and glucose-fed cells. Results showed that cytosolic free-histidine concentration was higher and more sensitive to the environment than free histidine in the mitochondria. Moreover, histidine was readily transported across the plasma membrane and mitochondrial inner membrane, which had almost similar transport rates and transport constants, and histidine transport was not influenced by cellular metabolic state. These sensors are potential tools for tracking histidine dynamics inside subcellular organelles, and they will open an avenue to explore complex histidine signaling.