Visualization of peroxynitrite-induced changes of labile Zn2+ in the endoplasmic reticulum with benzoresorufin-based fluorescent probes

Wei Lin, Daniela Buccella, Stephen J. Lippard

Research output: Contribution to journalArticle

Abstract

Zn2+ plays essential roles in biology, and the homeostasis of Zn2+ is tightly regulated in all cells. Subcellular distribution and trafficking of labile Zn2+, 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 Zn2+, 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 [N3O] binding motif that resembles that of previously reported fluorescein-based sensors of the Zinpyr family, reproducing well their binding capabilities and yielding comparable Kd values in the sub-nanomolar and picomolar ranges. The ZBR sensors display up to 8.4-fold emission fluorescence enhancement upon Zn2+ 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 2+ thus providing evidence for a direct connection between ER stress and ER Zn2+ homeostasis.

Original languageEnglish (US)
Pages (from-to)13512-13520
Number of pages9
JournalJournal of the American Chemical Society
Volume135
Issue number36
DOIs
StatePublished - Sep 11 2013

Fingerprint

Peroxynitrous Acid
Fluorescent Dyes
Endoplasmic Reticulum
Visualization
Zinc
Sensors
Homeostasis
Reactive Nitrogen Species
Endoplasmic Reticulum Stress
Neural Stem Cells
Optical Imaging
Fluorescein
Fluorescence Microscopy
Imaging techniques
Organelles
Amines
Fluorophores
Fluorescence microscopy
Fluorescence
Stem cells

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Visualization of peroxynitrite-induced changes of labile Zn2+ in the endoplasmic reticulum with benzoresorufin-based fluorescent probes. / Lin, Wei; Buccella, Daniela; Lippard, Stephen J.

In: Journal of the American Chemical Society, Vol. 135, No. 36, 11.09.2013, p. 13512-13520.

Research output: Contribution to journalArticle

@article{d215aa1481934622b6bc07899df84328,
title = "Visualization of peroxynitrite-induced changes of labile Zn2+ in the endoplasmic reticulum with benzoresorufin-based fluorescent probes",
abstract = "Zn2+ plays essential roles in biology, and the homeostasis of Zn2+ is tightly regulated in all cells. Subcellular distribution and trafficking of labile Zn2+, 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 Zn2+, 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 [N3O] binding motif that resembles that of previously reported fluorescein-based sensors of the Zinpyr family, reproducing well their binding capabilities and yielding comparable Kd values in the sub-nanomolar and picomolar ranges. The ZBR sensors display up to 8.4-fold emission fluorescence enhancement upon Zn2+ 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 2+ thus providing evidence for a direct connection between ER stress and ER Zn2+ homeostasis.",
author = "Wei Lin and Daniela Buccella and Lippard, {Stephen J.}",
year = "2013",
month = "9",
day = "11",
doi = "10.1021/ja4059487",
language = "English (US)",
volume = "135",
pages = "13512--13520",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "36",

}

TY - JOUR

T1 - Visualization of peroxynitrite-induced changes of labile Zn2+ in the endoplasmic reticulum with benzoresorufin-based fluorescent probes

AU - Lin, Wei

AU - Buccella, Daniela

AU - Lippard, Stephen J.

PY - 2013/9/11

Y1 - 2013/9/11

N2 - Zn2+ plays essential roles in biology, and the homeostasis of Zn2+ is tightly regulated in all cells. Subcellular distribution and trafficking of labile Zn2+, 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 Zn2+, 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 [N3O] binding motif that resembles that of previously reported fluorescein-based sensors of the Zinpyr family, reproducing well their binding capabilities and yielding comparable Kd values in the sub-nanomolar and picomolar ranges. The ZBR sensors display up to 8.4-fold emission fluorescence enhancement upon Zn2+ 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 2+ thus providing evidence for a direct connection between ER stress and ER Zn2+ homeostasis.

AB - Zn2+ plays essential roles in biology, and the homeostasis of Zn2+ is tightly regulated in all cells. Subcellular distribution and trafficking of labile Zn2+, 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 Zn2+, 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 [N3O] binding motif that resembles that of previously reported fluorescein-based sensors of the Zinpyr family, reproducing well their binding capabilities and yielding comparable Kd values in the sub-nanomolar and picomolar ranges. The ZBR sensors display up to 8.4-fold emission fluorescence enhancement upon Zn2+ 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 2+ thus providing evidence for a direct connection between ER stress and ER Zn2+ homeostasis.

UR - http://www.scopus.com/inward/record.url?scp=84884194583&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84884194583&partnerID=8YFLogxK

U2 - 10.1021/ja4059487

DO - 10.1021/ja4059487

M3 - Article

C2 - 23902285

AN - SCOPUS:84884194583

VL - 135

SP - 13512

EP - 13520

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 36

ER -