Development of Fluorogenic Reagent that Enables Simultaneous Detection and Labeling of Hydropersulfide.

Area of Science:

• Biochemistry
• Chemical Biology
• Proteomics

Background:

• Supersulfidation (S-sulfhydrylation) of cysteine residues (Cys-SH) significantly alters protein function and biological system activity.
• Existing methods for detecting Cys-SH modifications include fluorescence probes for real-time analysis and biotin tag-switch methods for proteome-wide labeling.
• A limitation of current techniques is the lack of a single system capable of both simultaneous detection and labeling of supersulfidated residues.

Purpose of the Study:

• To develop a novel system for the simultaneous real-time detection and labeling of cysteine persulfide (Cys-SSH).
• To create dinitrobenzene-based fluorogenic probes with tunable electrophilicity for selective reaction with supersulfides.

Main Methods:

• Design and synthesis of dinitrobenzene-based fluorogenic probes with adjustable electrophilicity.
• Utilizing the probes' reactivity with highly nucleophilic supersulfides like Cys-SSH and Na2S2.
• Simultaneous labeling of Cys-SSH with a dinitrobenzene tag during detection.

Main Results:

• The developed probes exhibit rapid and selective reactions with supersulfides.
• The probes enable simultaneous detection and labeling of Cys-SSH in a single system.
• This approach facilitates real-time analysis of supersulfidation events.

Conclusions:

• The novel dinitrobenzene probes offer a powerful tool for simultaneous supersulfide detection and labeling.
• This advancement provides new possibilities for studying the functional roles of supersulfidation in biological systems.
• The developed system addresses the need for integrated real-time analysis and labeling of modified cysteine residues.