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Live-Cell Imaging of Physiologically Relevant Metal Ions Using Genetically Encoded FRET-Based Probes

1
Gottfried Schatz Research Center, Chair of Molecular Biology and Biochemistry, Medical University of Graz, Neue Stiftingtalstraße 6/6, 8010 Graz, Austria
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BioTechMed Graz, Mozartgasse 12/II, 8010 Graz, Austria
*
Author to whom correspondence should be addressed.
Cells 2019, 8(5), 492; https://doi.org/10.3390/cells8050492
Received: 30 April 2019 / Revised: 17 May 2019 / Accepted: 21 May 2019 / Published: 22 May 2019
(This article belongs to the Special Issue Emerging Trends in Metal Biochemistry)
Essential biochemical reactions and processes within living organisms are coupled to subcellular fluctuations of metal ions. Disturbances in cellular metal ion homeostasis are frequently associated with pathological alterations, including neurotoxicity causing neurodegeneration, as well as metabolic disorders or cancer. Considering these important aspects of the cellular metal ion homeostasis in health and disease, measurements of subcellular ion signals are of broad scientific interest. The investigation of the cellular ion homeostasis using classical biochemical methods is quite difficult, often even not feasible or requires large cell numbers. Here, we report of genetically encoded fluorescent probes that enable the visualization of metal ion dynamics within individual living cells and their organelles with high temporal and spatial resolution. Generally, these probes consist of specific ion binding domains fused to fluorescent protein(s), altering their fluorescent properties upon ion binding. This review focuses on the functionality and potential of these genetically encoded fluorescent tools which enable monitoring (sub)cellular concentrations of alkali metals such as K+, alkaline earth metals including Mg2+ and Ca2+, and transition metals including Cu+/Cu2+ and Zn2+. Moreover, we discuss possible approaches for the development and application of novel metal ion biosensors for Fe2+/Fe3+, Mn2+ and Na+. View Full-Text
Keywords: FRET; fluorescent protein; FP; fluorescence microscopy; genetically encoded probes; imaging; Ca2+; Cu+; Cu2+; Fe2+; Fe3+; K+; Mg2+; Mn2+; Zn2+ FRET; fluorescent protein; FP; fluorescence microscopy; genetically encoded probes; imaging; Ca2+; Cu+; Cu2+; Fe2+; Fe3+; K+; Mg2+; Mn2+; Zn2+
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Bischof, H.; Burgstaller, S.; Waldeck-Weiermair, M.; Rauter, T.; Schinagl, M.; Ramadani-Muja, J.; Graier, W.F.; Malli, R. Live-Cell Imaging of Physiologically Relevant Metal Ions Using Genetically Encoded FRET-Based Probes. Cells 2019, 8, 492.

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