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Selective Labeling of Proteins on Living Cell Membranes Using Fluorescent Nanodiamond Probes

Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, Japan
PRESTO, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan
Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 615-8501, Japan
Department of Pediatrics, Graduate School of Medicine, Gifu University, Yanagido 1-1, Gifu 501-1194, Japan
Graduate School of Engineering, Osaka University, Yamadaoka 2-1, Suita, Osaka 565-0871, Japan
Immunology Frontier Research Center, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871, Japan
Department of Biophysics, Graduate School of Science, Kyoto University, Kitashirakawa-oiwake, Sakyo-ku, Kyoto 606-8502, Japan
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Yurii Gun'ko
Nanomaterials 2016, 6(4), 56;
Received: 22 December 2015 / Revised: 7 March 2016 / Accepted: 15 March 2016 / Published: 25 March 2016
(This article belongs to the Special Issue Nanoparticles in Bioimaging)
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. View Full-Text
Keywords: nanodiamond; nitrogen-vacancy center; polyglycerol; β-lactamase tag; membrane protein nanodiamond; nitrogen-vacancy center; polyglycerol; β-lactamase tag; membrane protein
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Sotoma, S.; Iimura, J.; Igarashi, R.; Hirosawa, K.M.; Ohnishi, H.; Mizukami, S.; Kikuchi, K.; Fujiwara, T.K.; Shirakawa, M.; Tochio, H. Selective Labeling of Proteins on Living Cell Membranes Using Fluorescent Nanodiamond Probes. Nanomaterials 2016, 6, 56.

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