Dynamic imaging of cytosolic zinc in Arabidopsis roots combining FRET sensors and RootChip technology
| Autor: | Wolf B. Frommer, Guido Grossmann, Jan L. Vinkenborg, Sébastien Thomine, Maarten Merkx, Viviane Lanquar |
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| Přispěvatelé: | Protein Engineering, Chemical Biology |
| Rok vydání: | 2013 |
| Předmět: |
Physiology
Microfluidics Arabidopsis Intracellular Space chemistry.chemical_element Plant Science Zinc Plant Roots Cofactor Cytosol Imaging Three-Dimensional Fluorescence Resonance Energy Transfer chemistry.chemical_classification biology biology.organism_classification Perfusion Enzyme Förster resonance energy transfer chemistry Biochemistry Biophysics biology.protein Extracellular Space Homeostasis |
| Zdroj: | New Phytologist, 202, 198-208. Wiley-Blackwell |
| ISSN: | 1469-8137 0028-646X |
| Popis: | Summary Zinc plays a central role in all living cells as a cofactor for enzymes and as a structural element enabling the adequate folding of proteins. In eukaryotic cells, metals are highly compartmentalized and chelated. Although essential to characterize the mechanisms of Zn2+ homeostasis, the measurement of free metal concentrations in living cells has proved challenging and the dynamics are difficult to determine. Our work combines the use of genetically encoded Forster resonance energy transfer (FRET) sensors and a novel microfluidic technology, the RootChip, to monitor the dynamics of cytosolic Zn2+ concentrations in Arabidopsis root cells. Our experiments provide estimates of cytosolic free Zn2+ concentrations in Arabidopsis root cells grown under sufficient (0.4 nM) and excess (2 nM) Zn2+ supply. In addition, monitoring the dynamics of cytosolic [Zn2+] in response to external supply suggests the involvement of high- and low-affinity uptake systems as well as release from internal stores. In this study, we demonstrate that the combination of genetically encoded FRET sensors and microfluidics provides an attractive tool to monitor the dynamics of cellular metal ion concentrations over a wide concentration range in root cells. |
| Databáze: | OpenAIRE |
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