Selective Labeling of Proteins on Living Cell Membranes Using Fluorescent Nanodiamond Probes.
Selective Labeling of Proteins on Living Cell Membranes Using Fluorescent Nanodiamond Probes.
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DOI：
10.3390/nano6040056
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发表时间：
2016-03-25
期刊：
影响因子：
--
通讯作者：
Tochio H
中科院分区：
文献类型：
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作者：
Sotoma S;Iimura J;Igarashi R;Hirosawa KM;Ohnishi H;Mizukami S;Kikuchi K;Fujiwara TK;Shirakawa M;Tochio H
荧光纳米金刚石 (FND) 无可挑剔的光稳定性是用于活细胞中蛋白质荧光成像的理想特性。然而，此类应用需要用 FND 对目标蛋白进行高度特异性标记。此外，未修饰的 FND 的表面往往会非特异性地吸附生物分子，这阻碍了 FND 可靠地靶向蛋白质。在这里，我们将 FND 的超支化聚甘油修饰与 β-内酰胺酶标签系统相结合，开发了一种对细胞中感兴趣的蛋白质进行选择性成像的策略。这些技术的结合使得能够用 FND 对白细胞介素 18 受体 α 链（一种膜受体）进行位点特异性标记，最终能够跟踪 FND 标记的蛋白质在膜表面上的扩散轨迹。
The impeccable photostability of fluorescent nanodiamonds (FNDs) is an ideal property for use in fluorescence imaging of proteins in living cells. However, such an application requires highly specific labeling of the target proteins with FNDs. Furthermore, the surface of unmodified FNDs tends to adsorb biomolecules nonspecifically, which hinders the reliable targeting of proteins with FNDs. Here, we combined hyperbranched polyglycerol modification of FNDs with the β-lactamase-tag system to develop a strategy for selective imaging of the protein of interest in cells. The combination of these techniques enabled site-specific labeling of Interleukin-18 receptor alpha chain, a membrane receptor, with FNDs, which eventually enabled tracking of the diffusion trajectory of FND-labeled proteins on the membrane surface.