title: Heavy metal-free 19F NMR probes for quantitative measurements of glutathione reductase activity using silica nanoparticles as a signal quencher.
abstract: For the quantitative assessment of the glutathione reductase (GR) activity with a (19)F 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 (19)F 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 (19)F NMR signals of perfluorinated dendrimers were recovered. The enzymatic activity of GR was determined from the increase of the magnitude of (19)F 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.
authors.affiliation: Department of Polymer Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Tanaka
authors.forename: Narufumi
authors.initials: N
authors.lastname: Kitamura
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Chujo
mesh_terms: D050091:Dendrimers / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D057075:Enzyme Assays; D005461:Fluorine / Q000737:chemistry; D005980:Glutathione Reductase / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019216:Metals, Heavy / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D050091:Dendrimers; D004220:Disulfides; D019216:Metals, Heavy; D005461:Fluorine; D012822:Silicon Dioxide; D005980:Glutathione Reductase