Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

Monitoring cytosolic and ER Zn2+ in stimulated breast cancer cells using genetically encoded FRET sensors

Hessels, Anne M., Taylor, Kathryn Mary ORCID: and Merkx, Maarten 2016. Monitoring cytosolic and ER Zn2+ in stimulated breast cancer cells using genetically encoded FRET sensors. Metallomics 8 , pp. 211-217. 10.1039/C5MT00257E

[thumbnail of Monitoring Cytosolic and ER Zn2+ in Stimulated Breast Cancer Cells using Genetically Encoded FRET Sensors.pdf]
PDF - Published Version
Available under License Creative Commons Attribution No Derivatives.

Download (1MB) | Preview


The Zn2+-specific ion channel ZIP7 has been implicated to play an important role in releasing Zn2+ from the ER. External stimulation of breast cancer cells has been proposed to induce phosphorylation of ZIP7 by CK2α, resulting in ZIP7-mediated Zn2+ release from the ER into the cytosol. Here, we examined whether changes in cytosolic and ER Zn2+ concentrations can be detected upon such external stimuli. Two previously developed FRET sensors for Zn2+, eZinCh-2 (Kd = 1 nM at pH 7.1) and eCALWY-4 (Kd = 0.63 nM at pH 7.1), were expressed in both the cytosol and the ER of wild-type MCF-7 and TamR cells. Treatment of MCF-7 and TamR cells with external Zn2+ and pyrithione, one of the previously used triggers, resulted in an immediate increase in free Zn2+ in both cytosol and ER, suggesting that Zn2+ was directly transferred across the cellular membranes by pyrithione. Cells treated with a second trigger, EGF/ionomycin, showed no changes in intracellular Zn2+ levels, neither in multicolor imaging experiments that allowed simultaneous imaging of cytosolic and ER Zn2+, nor in experiments in which cytosolic and ER Zn2+ were monitored separately. In contrast to previous work using small-molecule fluorescent dyes, these results indicate that EGF–ionomycin treatment does not result in significant changes in cytosolic Zn2+ levels as a result from Zn2+ release from the ER. These results underline the importance of using genetically encoded fluorescent sensors to complement and verify intracellular imaging experiments with synthetic fluorescent Zn2+ dyes.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Pharmacy
Publisher: Royal Society of Chemistry
ISSN: 1756-5901
Date of First Compliant Deposit: 30 March 2016
Date of Acceptance: 22 December 2015
Last Modified: 02 May 2023 19:24

Citation Data

Cited 20 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item Edit Item


Downloads per month over past year

View more statistics