Direct Comparison of a Genetically Encoded Sensor and Small Molecule Indicator: Implications for Quantification of Cytosolic Zn2+

作者：Qin Yan; Miranda Jose G; Stoddard Caitlin I; Dean Kevin M; Galati Domenico F; Palmer Amy E^*

来源：ACS Chemical Biology, 2013, 8(11): 2366-2371.

DOI：10.1021/cb4003859

Fluorescent sensors are powerful tools for visualizing and quantifying molecules and ions in living cells. A variety of small molecule and genetically encoded sensors have been developed for studying intracellular Zn2+ homeostasis and signaling, but no direct comparisons exist, making it challenging for researchers to identify the appropriate sensor for a given application. Here we directly compare the widely used small molecule probe FluoZin-3 and a genetically encoded sensor, ZapCY2. We demonstrate that, in contrast to FluoZin-3, ZapCY2 exhibits a well-defined cytosolic localization, provides estimates of Zn2+ concentration with little variability, does not perturb cytosolic Zn2+ levels, and exhibits rapid Zn2+ response dynamics. ZapCY2 was used to measure Zn2+ concentrations in 5 different cell types, revealing higher cytosolic Zn2+ levels in prostate cancer cells compared to normal prostate cells (although the total zinc is reduced in prostate cancer cells), suggesting distinct regulatory mechanisms.

• 出版日期2013-11