A Synthetic Fluorescent Mitochondria‐Targeted Sensor for Ratiometric Imaging of Calcium in Live Cells. (7th June 2019)

• Record Type: Journal Article
• Authors: Pendin, Diana; Norante, Rosa; De Nadai, Andrea; Gherardi, Gaia; Vajente, Nicola; Basso, Emy; Kaludercic, Nina; Mammucari, Cristina; Paradisi, Cristina; Pozzan, Tullio; Mattarei, Andrea
• Is Part Of: Angewandte Chemie. Volume 131:Number 29(2019)
• Page Start: 10022
• Page End: 10027
• Publication Date: 2019-06-07
• Subjects: Biologische Bildgebung -- Ca2+-Sensor -- Calcium -- Fluoreszenzsonden -- Mitochondrien; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.201902272
• Languages: English
• ISSNs: 0044-8249

Abstract

Abstract: Ca 2+ handling by mitochondria is crucial for cell life and the direct measure of mitochondrial Ca 2+ concentration in living cells is of pivotal interest. Genetically‐encoded indicators greatly facilitated this task, however they require demanding delivery procedures. On the other hand, existing mitochondria‐targeted synthetic Ca 2+ indicators are plagued by several drawbacks, for example, non‐specific localization, leakage, toxicity. Here we report the synthesis and characterization of a new fluorescent Ca 2+ sensor, named mt‐fura‐2, obtained by coupling two triphenylphosphonium cations to the molecular backbone of the ratiometric Ca 2+ indicator fura‐2. Mt‐fura‐2 binds Ca 2+ with a dissociation constant of ≈1.5 μm in vitro. When loaded in different cell types as acetoxymethyl ester, the probe shows proper mitochondrial localization and accurately measures matrix [Ca 2+ ] variations, proving its superiority over available dyes. We describe the synthesis, characterization and application of mt‐fura‐2 to cell types where the delivery of genetically‐encoded indicators is troublesome. Abstract : Überwachung von Calcium : Ein selektiver Mitochondrien‐gerichteter ratiometrischer fluoreszierender Ca 2+ ‐Sensor, mt‐fura‐2, wird durch Kupplung von zwei Triphenylphosphoniumkationen an die Ca 2+ ‐Sonde fura‐2 erhalten. Wird die Sonde in Form des Acetoxymethylesters in Zellen geladen, wird sie effizient in den Mitochondrien lokalisiert und misst präzise die Ca 2+ ‐Konzentrationen.