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Nanomaterials 2016, 6(4), 56; doi:10.3390/nano6040056

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

1
Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510, Japan
2
PRESTO, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan
3
Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 615-8501, Japan
4
Department of Pediatrics, Graduate School of Medicine, Gifu University, Yanagido 1-1, Gifu 501-1194, Japan
5
Graduate School of Engineering, Osaka University, Yamadaoka 2-1, Suita, Osaka 565-0871, Japan
6
Immunology Frontier Research Center, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871, Japan
7
Department of Biophysics, Graduate School of Science, Kyoto University, Kitashirakawa-oiwake, Sakyo-ku, Kyoto 606-8502, Japan
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editor: Yurii Gun'ko
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)
View Full-Text   |   Download PDF [1679 KB, uploaded 25 March 2016]   |  

Abstract

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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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

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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