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      Advances and current challenges in calcium signaling

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          Calcium signaling.

          Calcium ions (Ca(2+)) impact nearly every aspect of cellular life. This review examines the principles of Ca(2+) signaling, from changes in protein conformations driven by Ca(2+) to the mechanisms that control Ca(2+) levels in the cytoplasm and organelles. Also discussed is the highly localized nature of Ca(2+)-mediated signal transduction and its specific roles in excitability, exocytosis, motility, apoptosis, and transcription.
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            Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin.

            Important Ca2+ signals in the cytosol and organelles are often extremely localized and hard to measure. To overcome this problem we have constructed new fluorescent indicators for Ca2+ that are genetically encoded without cofactors and are targetable to specific intracellular locations. We have dubbed these fluorescent indicators 'cameleons'. They consist of tandem fusions of a blue- or cyan-emitting mutant of the green fluorescent protein (GFP), calmodulin, the calmodulin-binding peptide M13, and an enhanced green- or yellow-emitting GFP. Binding of Ca2+ makes calmodulin wrap around the M13 domain, increasing the fluorescence resonance energy transfer (FRET) between the flanking GFPs. Calmodulin mutations can tune the Ca2+ affinities to measure free Ca2+ concentrations in the range 10(-8) to 10(-2) M. We have visualized free Ca2+ dynamics in the cytosol, nucleus and endoplasmic reticulum of single HeLa cells transfected with complementary DNAs encoding chimaeras bearing appropriate localization signals. Ca2+ concentration in the endoplasmic reticulum of individual cells ranged from 60 to 400 microM at rest, and 1 to 50 microM after Ca2+ mobilization. FRET is also an indicator of the reversible intermolecular association of cyan-GFP-labelled calmodulin with yellow-GFP-labelled M13. Thus FRET between GFP mutants can monitor localized Ca2+ signals and protein heterodimerization in individual live cells.
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              Mitochondria as sensors and regulators of calcium signalling.

              During the past two decades calcium (Ca(2+)) accumulation in energized mitochondria has emerged as a biological process of utmost physiological relevance. Mitochondrial Ca(2+) uptake was shown to control intracellular Ca(2+) signalling, cell metabolism, cell survival and other cell-type specific functions by buffering cytosolic Ca(2+) levels and regulating mitochondrial effectors. Recently, the identity of mitochondrial Ca(2+) transporters has been revealed, opening new perspectives for investigation and molecular intervention.
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                Author and article information

                Journal
                New Phytologist
                New Phytol
                Wiley
                0028646X
                April 2018
                April 2018
                January 14 2018
                : 218
                : 2
                : 414-431
                Affiliations
                [1 ]Institut für Biologie und Biotechnologie der Pflanzen; Universität Münster; Schlossplatz 7/8 48149 Münster Germany
                [2 ]Department of Molecular Plant Physiology and Biophysics; University Würzburg; Julius-von-Sachs Platz 2 97082 Würzburg Germany
                [3 ]Lehrstuhl für Botanik; Technische Universität München; Am Hochanger 4 D-85354 Freising Germany
                [4 ]Department of Modeling of Biological Processes; COS Heidelberg/Bioquant; Heidelberg University; Im Neuenheimer Feld 267 69120 Heidelberg Germany
                [5 ]Institute of Genetics; Biocenter University of Munich (LMU); Großhaderner Straße 4 82152 Martinsried Germany
                [6 ]Department of Plant Biochemistry; Dahlem Center of Plant Sciences; Freie Universität Berlin; 14195 Berlin Germany
                [7 ]Department of Developmental Biology; Centre for Organismal Studies (COS); University of Heidelberg; Im Neuenheimer Feld 230 69120 Heidelberg Germany
                Article
                10.1111/nph.14966
                © 2018

                http://doi.wiley.com/10.1002/tdm_license_1.1

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