Engineered Protein-tag for Rapid Live-cell Fluorogenic Visualization of Proteins by Anionic Probes
Engineered Protein-tag for Rapid Live-cell Fluorogenic Visualization of Proteins by Anionic Probes
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DOI：
10.1246/cl.190875
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发表时间：
2020-03-01
影响因子：
1.6
通讯作者：
Kikuchi, Kazuya
中科院分区：
文献类型：
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作者：
Gao, Jingchi;Hori, Yuichiro;Kikuchi, Kazuya
使用蛋白质标签和荧光探针进行活细胞蛋白质标记是研究蛋白质定位和动力学的有效方法。在这里，我们设计了一种蛋白质标签，PYP-标签，以高对比度对膜蛋白进行成像。 PYP-标签表面带电氨基酸的突变导致标记动力学和亮度的增强。此外，表面电荷的修饰并没有影响其在细胞表面的表达，我们成功地将这种标记技术应用于葡萄糖转运蛋白4易位的高对比度成像。
Live-cell protein labeling using a protein tag and a fluorescent probe is a powerful approach for studying protein localization and dynamics. Herein, we engineered a protein tag, PYP-tag, to image membrane proteins with high contrast. Mutations of charged amino acids on the surface of PYP-tag led to the enhancement of labeling kinetics and brightness. Furthermore, the modification of the surface charge did not affect its expression on the cell surface, and we succeeded in applying this labeling technique to high-contrast imaging of translocation of glucose transporter 4.