A highly selective and sensitive fluorescence sensing system for distinction between diphosphate and nucleoside...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Molecular Biology

Background:

• Distinguishing between diphosphate (PPi) and nucleoside triphosphates (NTPs) is crucial for various bioanalytical applications.
• Existing chemosensors often lack selectivity, limiting their practical use in complex biological samples.

Purpose of the Study:

• To develop a novel fluorescence sensing system capable of selectively differentiating PPi from NTPs.
• To enable sensitive detection and quantification of PPi at nanomolar concentrations.
• To apply the developed sensing system for direct quantification of RNA polymerase activity.

Main Methods:

• Development of a dual-sensor system combining a binuclear Zn(II) complex (1·2Zn) and boronic acid (BA).
• Utilizing differential signal responses of the two sensors to PPi and NTPs for selective detection.
• Employing fluorescence spectroscopy for signal monitoring and quantification.

Main Results:

• The combined sensing system successfully distinguishes PPi from NTPs.
• The system demonstrates high sensitivity, detecting PPi at nanomolar concentrations.
• The sensing system was effectively applied to directly quantify RNA polymerase activity.

Conclusions:

• A novel and selective fluorescence sensing system for PPi detection has been established.
• This system overcomes limitations of existing chemosensors, enabling new bioanalytical applications.
• The developed method provides a sensitive tool for enzyme activity assays, such as RNA polymerase quantification.