Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes
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...
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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American Chemical Society (ACS)
2014
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| Online Access: | http://hdl.handle.net/1721.1/88575 https://orcid.org/0000-0002-2693-4982 |
| _version_ | 1826198908650389504 |
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| author | Lin, Wei Buccella, Daniela Lippard, Stephen J. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Lin, Wei Buccella, Daniela Lippard, Stephen J. |
| author_sort | Lin, Wei |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:11:51Z |
| format | Article |
| id | mit-1721.1/88575 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:11:51Z |
| publishDate | 2014 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/885752022-10-01T01:57:18Z Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes Lin, Wei Buccella, Daniela Lippard, Stephen J. Massachusetts Institute of Technology. Department of Chemistry Lippard, Stephen J. Lin, Wei Buccella, Daniela Lippard, Stephen J. 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. National Institute of General Medical Sciences (U.S.) (Grant GM065519) 2014-08-07T14:37:32Z 2014-08-07T14:37:32Z 2013-07 2013-06 Article http://purl.org/eprint/type/JournalArticle 0002-7863 1520-5126 http://hdl.handle.net/1721.1/88575 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. https://orcid.org/0000-0002-2693-4982 en_US http://dx.doi.org/10.1021/ja4059487 Journal of the American Chemical Society Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society (ACS) Prof. Lippard via Erja Kajosalo |
| spellingShingle | Lin, Wei Buccella, Daniela Lippard, Stephen J. Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes |
| title | Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes |
| title_full | Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes |
| title_fullStr | Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes |
| title_full_unstemmed | Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes |
| title_short | Visualization of Peroxynitrite-Induced Changes of Labile Zn[superscript 2+] in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes |
| title_sort | visualization of peroxynitrite induced changes of labile zn superscript 2 in the endoplasmic reticulum with benzoresorufin based fluorescent probes |
| url | http://hdl.handle.net/1721.1/88575 https://orcid.org/0000-0002-2693-4982 |
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