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A Synthetic Fluorescent Mitochondria‐Targeted Sensor for Ratiometric Imaging of Calcium in Live Cells
Author(s) -
Pendin Diana,
Norante Rosa,
De Nadai Andrea,
Gherardi Gaia,
Vajente Nicola,
Basso Emy,
Kaludercic Nina,
Mammucari Cristina,
Paradisi Cristina,
Pozzan Tullio,
Mattarei Andrea
Publication year - 2019
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201902272
Subject(s) - fluorescence , mitochondrion , fura 2 , cell , chemistry , biophysics , in vitro , calcium , cytosol , microbiology and biotechnology , biochemistry , biology , enzyme , physics , organic chemistry , quantum mechanics
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 μ m 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.