title: Development of novel FP-based probes for live-cell imaging of nitric oxide dynamics.
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 Ca(2+) sensor allowed us to visualize and Ca(2+) 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.
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Emrah
authors: E
authors: Eroglu
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Benjamin
authors: B
authors: Gottschalk
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Suphachai
authors: S
authors: Charoensin
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Sandra
authors: S
authors: Blass
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Helmut
authors: H
authors: Bischof
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Rene
authors: R
authors: Rost
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Corina T
authors: CT
authors: Madreiter-Sokolowski
authors: Institute of Biophysics, Center of Physiological Medicine, Medical University of Graz, Harrachgasse 21/IV, 8010 Graz, Austria.
authors: Brigitte
authors: B
authors: Pelzmann
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Eva
authors: E
authors: Bernhart
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Wolfgang
authors: W
authors: Sattler
authors: Institute of Physiological Chemistry, Center of Physiological Medicine, Medical University of Graz, Harrachgasse 21/II, 8010 Graz, Austria.
authors: Seth
authors: S
authors: Hallström
authors: Nanomedical Research Laboratory, Department of Chemistry and Biochemistry, Ohio University, 350 West State Street, Athens, Ohio 45701, USA.
authors: Tadeusz
authors: T
authors: Malinski
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Markus
authors: M
authors: Waldeck-Weiermair
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Wolfgang F
authors: WF
authors: Graier
authors: Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
authors: Roland
authors: R
authors: Malli
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D020013:Calcium Signaling; D002642:Chick Embryo; D042783:Endothelial Cells / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006352:Heart Ventricles / Q000166:cytology; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C527701:enhanced cyan fluorescent protein; C485192:green fluorescent protein, Aequorea victoria; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D009569:Nitric Oxide