Benzylic Trifluoromethyl Accelerates 1,6-Elimination Toward Rapid Probe Activation

Area of Science:

• Chemical Biology
• Biochemistry
• Cellular Imaging

Background:

• Hydrogen sulfide (H2S) is a crucial signaling molecule with diverse physiological roles.
• Accurate detection of intracellular H2S is essential for understanding its reactivity and effects.
• Existing methods for H2S detection often lack speed and sensitivity.

Purpose of the Study:

• To develop a highly efficient fluorescent probe for activity-based detection of hydrogen sulfide in live cells.
• To investigate the mechanism of probe activation and its dependence on H2S.
• To compare the performance of the novel probe with existing probes for H2S detection.

Main Methods:

• Design and synthesis of a novel resorufin-based fluorescent probe incorporating an α-CF3-benzyl group.
• Investigation of H2S-triggered 1,6-elimination reaction mechanism using computational chemistry.
• Evaluation of probe performance in live HeLa cells, assessing signal generation speed and sensitivity.

Main Results:

• A novel fluorescent probe, 4-azido-(α-CF3)-benzyl resorufin, was developed for H2S detection.
• The probe exhibits significantly faster signal generation and improved sensitivity compared to non-fluorinated analogs.
• Computational studies confirmed that the CF3 group lowers the activation energy barrier for probe activation.

Conclusions:

• The developed probe enables near-real-time detection of intracellular H2S.
• This advancement facilitates a deeper understanding of H2S reactivity and its physiological implications.
• The novel probe design represents a significant improvement for cellular H2S imaging.