Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes
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Lippard_Visualization.pdf
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Author(s)
Lin, Wei
Buccella, Daniela
Lippard, Stephen J.
Date Issued
July 2013
Journal
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Citation
Lin, Wei, Daniela Buccella, and Stephen J. Lippard. “Visualization of Peroxynitrite-Induced Changes of Labile Zn2+ in the Endoplasmic Reticulum with Benzoresorufin-Based Fluorescent Probes.” Journal of the American Chemical Society 135, no. 36 (September 11, 2013): 13512–13520.
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Author's final manuscript
Abstract
Zn[superscript 2+] plays essential roles in biology, and the homeostasis of Zn[superscript 2+] is tightly regulated in all cells. Subcellular distribution and trafficking of labile Zn[superscript 2+], and its inter-relation with reactive nitrogen species, are poorly understood due to the scarcity of appropriate imaging tools. We report a new family of red-emitting fluorescent sensors for labile Zn[superscript 2+], ZBR1–3, based on a benzoresorufin platform functionalized with dipicolylamine or picolylamine-derived metal binding groups. In combination, the pendant amines and fluorophore afford an [N[subscript 3]O] binding motif that resembles that of previously reported fluorescein-based sensors of the Zinpyr family, reproducing well their binding capabilities and yielding comparable K[subscript d] values in the sub-nanomolar and picomolar ranges. The ZBR sensors display up to 8.4-fold emission fluorescence enhancement upon Zn[superscript 2+] binding in the cuvette, with similar responses obtained in live cells using standard wide-field fluorescence microscopy imaging. The new sensors localize spontaneously in the endoplasmic reticulum (ER) of various tested cell lines, allowing for organelle-specific monitoring of zinc levels in live cells. Study of ER zinc levels in neural stem cells treated with a peroxynitrite generator, Sin-1, revealed an immediate decrease in labile Zn[superscript 2+] thus providing evidence for a direct connection between ER stress and ER Zn[superscript 2+] homeostasis.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
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DOI of Published Version
https://doi.org/10.1021/ja4059487
