Green Fluorescent Protein GFP-Chromophore-Based Probe for the Detection of Mitochondrial Viscosity in Living Cells
1The Education Ministry Key Laboratory of Resource Chemistry, Shanghai Key Laboratory of Rare Earth Functional Materials, and Shanghai Municipal Education Committee Key Laboratory of Molecular Imaging Probes and Sensors, Department of Chemistry, Shanghai Normal University, 100 Guilin Road, Shanghai 200234, China.
Abstract:
Viscosity is a pivotal factor for indicating the dysfunction of the mitochondria. To date, most of the fluorescent probes developed for mitochondrial viscosity have been designed using BODIPY, hemicyanine, or pyridine-based molecular rotors as part of the core structure. Our aim with this research was to extend the range of suitable fluorophores available for the construction of such fluorescent molecular rotors for evaluating the viscosity of mitocondria. Herein, we have developed a green fluorescent protein (GFP)-chromophore-based fluorescent probe (MIT-V) for the detection of mitochondrial viscosity in live cells. MIT-V exhibited a high sensitivity toward viscosity (from 7.9 cP to 438.4 cP). The "off-on" sensing mechanism of MIT-V was ascribed to the restricted rotation of single bonds and excited-state C═C double bonds of MIT-V. Cell studies indicated that MIT-V targets the mitochondria and that it was able to monitor real-time changes in the viscosity of live HeLa cell mitochondria. Therefore, we propose that MIT-V can be used as an effective chemosensor for the real-time imaging of mitochondrial viscosity in live cells. Our results clearly demonstrate the utility of such GFP-chromophore-based derivatives for the development of viscosity-sensitive systems.
Insights
Researchers developed a novel fluorescent probe, MIT-V, for detecting mitochondrial viscosity in live cells. This probe, based on a green fluorescent protein (GFP)-chromophore, offers a new tool for real-time monitoring of mitochondrial health.
Area of Science:
- Biochemistry
- Cell Biology
- Biophysics
Background:
- Mitochondrial viscosity is a key indicator of cellular dysfunction.
- Existing fluorescent probes for mitochondrial viscosity often rely on BODIPY, hemicyanine, or pyridine cores.
- There is a need for novel fluorophores to develop advanced viscosity-sensing systems.
Purpose of the Study:
- To develop a new class of fluorescent probes for evaluating mitochondrial viscosity.
- To create a green fluorescent protein (GFP)-chromophore-based probe for live-cell imaging of mitochondrial viscosity.
- To investigate the sensing mechanism and performance of the developed probe.
Main Methods:
- Synthesis of a novel GFP-chromophore-based fluorescent probe, MIT-V.
- Characterization of MIT-V's sensitivity to viscosity changes (7.9 cP to 438.4 cP).
- Live-cell imaging studies using HeLa cells to assess mitochondrial targeting and real-time viscosity monitoring.
Main Results:
- MIT-V successfully targets mitochondria in live cells.
- The probe exhibits high sensitivity to a wide range of viscosity.
- An "off-on" sensing mechanism was observed, attributed to restricted bond rotation within the probe.
- Real-time changes in mitochondrial viscosity were effectively monitored in live HeLa cells.
Conclusions:
- MIT-V is a novel and effective chemosensor for real-time imaging of mitochondrial viscosity.
- The developed GFP-chromophore-based probe expands the toolkit for viscosity-sensitive systems.
- This research highlights the potential of GFP-chromophore derivatives for creating advanced biological probes.
