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Chloro-Functionalized Photo-crosslinking BODIPY for Glutathione Sensing and Subcellular Trafficking
Dhiraj P Murale1, Seong Cheol Hong1,2, Md Mamunul Haque1
1Molecular Recognition Research Center, Korea Institute of Science and Technology, 5 Hwarang-ro 14gil, Seongbuk-gu, Seoul, 02792, Republic of Korea.
Chembiochem : a European Journal of Chemical Biology
|March 15, 2018
Summary
Researchers developed a new fluorescent probe to visualize glutathione (GSH) and its complex with glutathione S-transferase (GST) in live cells. This probe successfully tracked GSH translocation into mitochondria, offering new insights into cellular antioxidant responses.
Area of Science:
- Cell Biology
- Biochemistry
- Fluorescent Probes
Background:
- Glutathione (GSH) is a crucial intracellular antioxidant regulating cellular redox homeostasis.
- Existing probes for visualizing GSH in live cells lack detailed cellular localization information.
- Developing novel probes is essential for understanding GSH dynamics and cellular responses.
Purpose of the Study:
- To develop a novel fluorescent probe for in situ visualization of GSH in live cells.
- To investigate the cellular localization and dynamics of GSH, particularly its translocation.
- To image the glutathione S-transferase (GST)-GSH complex within live cells.
Main Methods:
- Synthesis and characterization of a new cell-permeable fluorescent probe (pcBD2-Cl).
- In situ labeling of GSH to form pcBD2-GSH, which exhibits fluorescence quenching and recovery upon GST binding.
- Photo-crosslinking assays to confirm probe-GST interactions under live-cell conditions.
- Co-staining with MitoTracker and mitochondrial fractionation to determine subcellular localization of pcBD2-GSH.
Main Results:
- The developed probe selectively reacts with GSH in situ.
- Fluorescence is quenched upon labeling but recovered upon binding to cellular glutathione S-transferase (GST), enabling imaging of the GSH-GST complex.
- Probe-GST interactions were confirmed via photo-crosslinking in live cells.
- Experiments demonstrated that pcBD2-GSH accumulates in mitochondria, indicating GSH translocation from the cytosol.
Conclusions:
- A novel fluorescent probe (pcBD2-Cl) was successfully developed for live-cell imaging of GSH and its complex with GST.
- This probe enables visualization of GSH translocation into mitochondria, a previously unobserved phenomenon.
- This work provides a new tool for studying cellular redox homeostasis and mitochondrial function.
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