Title

Optical Recording of Zn2+ Dynamics in the Mitochondrial Matrix and Intermembrane Space with the GZnP2 Sensor

Document Type

Article

Publication Date

6-20-2018

Keywords

Zinc ion, Mitochondrial matrix, Intermembrane space, Fluorescent sensor GZnP1

Organizational Units

College of Natual Science and Mathematics, Biological Sciences

Abstract

The zinc ion (Zn2+) is emerging as an important signaling molecule. Here, we engineered an improved Zn2+ probe GZnP2 based on a previously developed fluorescent sensor GZnP1 to provide a higher fluorescent readout (2-fold higher) that is proportional to cellular labile Zn2+ concentrations. We further developed a set of GZnP2 derived imaging tools to determine the labile Zn2+ concentrations in the mitochondrial matrix, mitochondrial intermembrane space (IMS), and cytosol in four different cell lines (HeLa, Cos-7, HEK293, and INS-1). The labile Zn2+ concentration in the matrix was less than 1 pM, while the labile Zn2+ concentration in the IMS was comparable to the cytosol (∼100 pM). With these sensors, we showed that upon exposure to high Zn2+, only the cytosol and the IMS were overloaded with Zn2+, while the mitochondrial matrix was unable to sequester excess labile Zn2+ in depolarized INS-1 cells. This work highlighted the importance of distinguishing the labile Zn2+ concentrations and dynamics between the mitochondrial matrix and IMS.

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