Multi-state fluorescent response system based on metal-induced protein phase transition and its application in skin barrier

By constructing a multi-state fluorescence response system based on metal-induced protein phase transitions, and utilizing the interaction between Hg2+ and CoA with H39-GFP, the problem of high selectivity and high sensitivity in existing ChAT detection methods in the skin barrier environment is solved, enabling real-time, dynamic monitoring and evaluation of ChAT activity.

CN122104864APending Publication Date: 2026-05-29NINGBO COLLEGE OF HEALTH SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NINGBO COLLEGE OF HEALTH SCI
Filing Date
2026-01-12
Publication Date
2026-05-29

AI Technical Summary

Technical Problem

Existing ChAT detection methods are difficult to achieve highly selective and sensitive in situ, real-time or dynamic analysis in the skin barrier environment, and sample processing is complex and difficult to adapt to complex biological barrier environments.

Method used

A multi-state fluorescence response system based on metal-induced protein phase transitions was constructed. The fluorescence signal was regulated by the interaction between Hg2+, CoA and supercharged green fluorescent protein (H39-GFP) and used to detect the activity of choline acetyltransferase (ChAT).

Benefits of technology

This invention provides a highly sensitive, specific, easy-to-operate, and low-cost fluorescence response system that can monitor changes in skin barrier function and ChAT activity in real time. It is suitable for various environmental conditions and fills the gap in dynamic evaluation technology.

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Abstract

The application provides a multi-state fluorescence response system based on metal-induced protein phase transition and application thereof in skin barrier. By exploring the interaction between CoA, Hg(II) and H39-GFP, a fluorescence response mechanism dependent on the change of metal ion concentration is constructed. Coenzyme A competes with Hg(II) through its thiol group, reduces the concentration of free Hg(II), and reduces the quenching effect of H39-GFP fluorescence. The system can sensitively detect choline acetyltransferase (ChAT) activity and quantitatively evaluate its expression level through the change of fluorescence intensity. HaCaT cells are used as experimental models to verify the application of the system in skin barrier research. Experiments show that the skin barrier function is significantly regulated by cell density, environmental factors (such as SLS, ultraviolet, calcium ion concentration), which provides new experimental basis for skin damage repair and related disease treatment. The fluorescence response system provides a new tool with high sensitivity for real-time monitoring of skin barrier function and related enzyme activity.
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