Exploring dynamics of molybdate in living animal cells by a genetically encoded FRET nanosensor
| Autor: | Tomoaki Niimi, Rie Tomioka, Hanayo Ueoka-Nakanishi, Syuntaro Iida, Yoichi Nakanishi, Masayoshi Maeshima |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Time Factors
Bone and Mineral Metabolism Green Fluorescent Proteins Sulfur metabolism Biophysics lcsh:Medicine Oxyanion Biosensing Techniques Molybdate Redox Biochemistry Protein Chemistry Sensitivity and Specificity Transmembrane Transport Proteins Diffusion chemistry.chemical_compound Bacterial Proteins In vivo Fluorescence Resonance Energy Transfer Humans Nanotechnology lcsh:Science Biology Bioinorganic Chemistry Molybdenum Oxalates Multidisciplinary Cofactors HEK 293 cells lcsh:R Proteins Biological Transport Cytosol Luminescent Proteins Förster resonance energy transfer Metabolism HEK293 Cells chemistry Bionanotechnology lcsh:Q Peptides Oxidation-Reduction Research Article Biotechnology Transcription Factors |
| Zdroj: | PLoS ONE, Vol 8, Iss 3, p e58175 (2013) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Molybdenum (Mo) is an essential trace element for almost all living organisms including animals. Mo is used as a catalytic center of molybdo-enzymes for oxidation/reduction reactions of carbon, nitrogen, and sulfur metabolism. Whilst living cells are known to import inorganic molybdate oxyanion from the surrounding environment, the in vivo dynamics of cytosolic molybdate remain poorly understood as no appropriate indicator is available for this trace anion. We here describe a genetically encoded Forester-resonance-energy-transfer (FRET)-based nanosensor composed of CFP, YFP and the bacterial molybdate-sensor protein ModE. The nanosensor MolyProbe containing an optimized peptide-linker responded to nanomolar-range molybdate selectively, and increased YFP:CFP fluorescence intensity ratio by up to 109%. By introduction of the nanosensor, we have been able to successfully demonstrate the real-time dynamics of molybdate in living animal cells. Furthermore, time course analyses of the dynamics suggest that novel oxalate-sensitive- and sulfate-resistant- transporter(s) uptake molybdate in a model culture cell. |
| Databáze: | OpenAIRE |
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