Development of novel FP-based probes for live-cell imaging of nitric oxide dynamics

Emrah Eroglu, Benjamin Gottschalk, Suphachai Charoensin, Sandra Blass, Helmut Bischof, Rene Rost, Corina T. Madreiter-Sokolowski, Brigitte Pelzmann, Eva Bernhart, Wolfgang Sattler, Seth Hallström, Tadeusz Malinski, Markus Waldeck-Weiermair, Wolfgang F. Graier and Roland Malli ()
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Emrah Eroglu: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Benjamin Gottschalk: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Suphachai Charoensin: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Sandra Blass: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Helmut Bischof: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Rene Rost: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Corina T. Madreiter-Sokolowski: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Brigitte Pelzmann: Institute of Biophysics, Center of Physiological Medicine, Medical University of Graz
Eva Bernhart: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Wolfgang Sattler: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Seth Hallström: Institute of Physiological Chemistry, Center of Physiological Medicine, Medical University of Graz
Tadeusz Malinski: Nanomedical Research Laboratory, Ohio University
Markus Waldeck-Weiermair: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Wolfgang F. Graier: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz
Roland Malli: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz

Nature Communications, 2016, vol. 7, issue 1, 1-11

Abstract: Abstract Nitric oxide ( ) is a free radical with a wide range of biological effects, but practically impossible to visualize in single cells. Here we report the development of novel multicoloured fluorescent quenching-based probes by fusing a bacteria-derived -binding domain close to distinct fluorescent protein variants. These genetically encoded probes, referred to as geNOps, provide a selective, specific and real-time read-out of cellular dynamics and, hence, open a new era of bioimaging. The combination of geNOps with a Ca2+ sensor allowed us to visualize and Ca2+ signals simultaneously in single endothelial cells. Moreover, targeting of the probes was used to detect signals within mitochondria. The geNOps are useful new tools to further investigate and understand the complex patterns of signalling on the single (sub)cellular level.

Date: 2016
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DOI: 10.1038/ncomms10623

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