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Ternary Zn(II) Complexes of Fluorescent Zinc Probes Zinpyr-1 and Zinbo-5 with the Low Molecular Weight Component of Exchangeable Cellular Zinc Pool
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    Ternary Zn(II) Complexes of Fluorescent Zinc Probes Zinpyr-1 and Zinbo-5 with the Low Molecular Weight Component of Exchangeable Cellular Zinc Pool
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    • Ilona Marszałek
      Ilona Marszałek
      Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland
    • Wojciech Goch
      Wojciech Goch
      Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland
      Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-091 Warsaw, Poland
    • Wojciech Bal*
      Wojciech Bal
      Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland
      *E-mail: [email protected]
      More by Wojciech Bal
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    Inorganic Chemistry

    Cite this: Inorg. Chem. 2019, 58, 21, 14741–14751
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    https://doi.org/10.1021/acs.inorgchem.9b02419
    Published October 24, 2019
    Copyright © 2019 American Chemical Society

    Abstract

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    The intracellular exchangeable Zn(II) is usually measured with synthetic fluorescent zinc sensors. 4′,5′-Bis[bis(2-pyridylmethyl)aminomethyl]-2′,7′-dichlorofluorescein (Zinpyr-1) is a sensor containing the fluorescein platform and a duplicated chelating unit. Its advantages include brightness and a relatively high affinity for Zn(II), Kd = 0.7 nM. 2-(4,5-Dimethoxy-2-hydroxyphenyl)-4-(2-pyridylmethyl)aminomethylbenzoxazole (Zinbo-5) is a member of a growing family of ratiometric synthetic Zn(II) probes, offering a possibility to determine Zn(II) concentration independently of the sensor concentration. Cells, however, contain high, millimolar or nearly millimolar concentrations of low molecular weight ligands (LMWLs) capable of binding Zn(II) ions. Previously, we demonstrated that such LMWLs can perturb the performance of some fluorescent zinc sensors by competition and formation of ternary Zn(sensor) (LMWL) complexes. Here we tested Zinpyr-1 and Zinbo-5 in this respect. Despite structural differences, both sensors formed such ternary complexes. We determined their stability constants CKtern and performed numerical simulations of Zn(II) distributions at physiological concentrations of selected LMWLs. Glutamic acid was found to provide the strongest ternary complexes with either of the studied sensors. Zn(Zinpyr-1)(Glu) was an absolutely dominant Zn(II)/Zinpyr-1 species (more than 96% of the exchangeable Zn(II)), and Zn(Zinbo-5)(Glu) was the most abundant one (more than 40%) in these simulations. Our results indicate that under cellular conditions these sensors are able to report Zn(II) complexed to LMWLs rather than free Zn2+ ions. On the other hand, the specific affinity of Zn(Zinpyr-1) and Zn(Zinbo-5) for Glu creates interesting opportunities for determining glutamic acid in biological samples.

    Copyright © 2019 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.inorgchem.9b02419.

    • All fluorescence spectra, full species distribution diagrams, the UV–vis spectra of Zn(Zinbo-5) titrated with GSH, and all numerical simulation results (PDF)

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    Cited By

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    This article is cited by 11 publications.

    1. Marek Łuczkowski, Weronika Leszczyńska, Joanna Wątły, Stephan Clemens, Artur Krężel. Phytochelatins Bind Zn(II) with Micro- to Picomolar Affinities without the Formation of Binuclear Complexes, Exhibiting Zinc Buffering and Muffling Rather than Storing Functions. Inorganic Chemistry 2024, 63 (24) , 10915-10931. https://doi.org/10.1021/acs.inorgchem.4c01707
    2. Claire M. S. Michielsen, Eva A. van Aalen, Maarten Merkx. Ratiometric Bioluminescent Zinc Sensor Proteins to Quantify Serum and Intracellular Free Zn2+. ACS Chemical Biology 2022, 17 (6) , 1567-1576. https://doi.org/10.1021/acschembio.2c00227
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    Inorganic Chemistry

    Cite this: Inorg. Chem. 2019, 58, 21, 14741–14751
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acs.inorgchem.9b02419
    Published October 24, 2019
    Copyright © 2019 American Chemical Society

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