Related Experiment Video
Updated: Jun 16, 2026
08:42
Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm
Published on: October 4, 2018
Dynamic copper(I) imaging in mammalian cells with a genetically encoded fluorescent copper(I) sensor
Seraphine V Wegner1, Hasan Arslan, Murat Sunbul
1Department of Chemistry and Institute for Biophysical Dynamics, The University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.
Journal of the American Chemical Society
|February 6, 2010
Summary
Researchers developed a novel fluorescent sensor, Amt1-FRET, to visualize cellular copper availability. This tool accurately measures low copper levels, crucial for understanding cellular processes and toxicity.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Imaging
Background:
- Copper is essential for life but toxic at high concentrations, necessitating strict cellular control.
- Visualizing intracellular copper availability is challenging due to its tight binding environment.
Purpose of the Study:
- To develop a genetically encoded fluorescent sensor for visualizing cellular copper(I) availability.
- To characterize the sensor's sensitivity, selectivity, and dynamic range in different cell types.
Main Methods:
- Engineered a fluorescent sensor (Amt1-FRET) based on the copper-responsive transcriptional regulator Amt1.
- Utilized Förster Resonance Energy Transfer (FRET) to detect copper(I)-induced conformational changes.
- Assessed sensor performance in yeast and mammalian cells.
Main Results:
- Amt1-FRET is a ratiometric sensor with extremely high affinity for copper(I) (K(d) = 2.5 x 10(-18) M).
- The sensor confirmed very low copper availability in yeast, reflecting the upper limit of cellular copper.
- Amt1-FRET operates within the cellular copper buffer range in mammalian cells, reporting dynamic copper fluctuations within minutes.
Conclusions:
- Amt1-FRET provides a sensitive and selective method for visualizing intracellular copper(I) availability.
- The sensor is valuable for studying copper homeostasis and dynamics in various biological systems.
- This tool aids in understanding the tight control of copper levels in cells.
