期刊
BIOORGANIC & MEDICINAL CHEMISTRY
卷 20, 期 1, 页码 96-100出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.bmc.2011.11.026
关键词
Glutathione reductase; F-19 NMR; Silica nanoparticle; Molecular probe; Quantitative assay
资金
- Magnetic Health Science Foundation
- Grants-in-Aid for Scientific Research [22655033, 22750107] Funding Source: KAKEN
For the quantitative assessment of the glutathione reductase (GR) activity with a F-19 NMR spectroscopy, we developed the heavy metal-free probes based on silica nanoparticles modified with water-soluble perfluorinated dendrimers via the disulfide linkers. Before enzymatic reaction, the molecular rotation of the perfluorinated dendrimers is highly restricted, and the magnitude of F-19 NMR signals from the perfluorinated dendrimers can be suppressed. By the reductive cleavage of the disulfide linkers with the reduced glutathione-mediated enzymatic reaction of GR, perfluorinated dendrimers can be released from the surfaces of the nanoparticles. Consequently, the F-19 NMR signals of perfluorinated dendrimers were recovered. The enzymatic activity of GR was determined from the increase of the magnitude of F-19 NMR signals. Finally, to demonstrate the feasibility of the probe in the presence of miscellaneous molecules under bio-mimetic conditions, the comparison study was executed with the cancer cell lysate. The value determined from our method showed a good agreement with that from the conventional method. (C) 2011 Elsevier Ltd. All rights reserved.
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