Fluorescence-plane polarization for the real-time monitoring of transferase migration in living cells

Yafu Wang, Huiyu Niu, Kui Wang, Liu Yang, Ge Wang, Tony D. James, Jiangli Fan, Hua Zhang

Research output: Contribution to journalArticlepeer-review

4 Citations (SciVal)

Abstract

Transferases are enzymes that exhibit multisite migration characteristics. Significantly, enzyme activity undergoes changes during this migration process, which inevitably impacts the physiological function of living organisms and can even lead to related malignant diseases. However, research in this field has been severely hindered by the lack of tools for the simultaneous and differential monitoring of site-specific transferase activity. Herein, we propose a novel strategy that integrates a fluorescence signal response with high sensitivity and an optical rotation signal response with superior spatial resolution. To validate the feasibility of this strategy, transferase γ-glutamyltransferase (GGT) was used as a model system to develop dual-mode chiral probes ACx-GGTB (AC17-GGTB and AC15-GGTB) using chiral amino acids as specific bifunctional recognition groups. The probes undergo structural changes under GGT, resulting in the release of bifunctional recognition groups (chiral amino acids) and simultaneously generate fluorescence signals and optical rotation signals. This dual-mode output exhibits high sensitivity and facilitates differentiation of sites. Furthermore, it enables simultaneous and differential detection of GGT activity at different sites during migration. We anticipate that probes developed based on this strategy will facilitate imaging-based monitoring of the activity for other transferases, thus providing an imaging platform suitable for the real-time tracking of transferase activity changes during migration.

Original languageEnglish
Pages (from-to)16291-16299
Number of pages9
JournalChemical Science
Volume15
Issue number39
Early online date12 Sept 2024
DOIs
Publication statusPublished - 21 Oct 2024

Data Availability Statement

The data supporting this article have been included as part of the ESI.†

Funding

This work was supported by the National Natural Science Foundation of China (U21A20314, 21925802, 22107089, and 52271251); the Program for Innovative Research Team in Science and Technology in University of Henan Province (23IRTSTHN002); T. D. J. wishes to thank the Royal Society for a Wolfson Research Merit Award and the Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University for support (2020ZD01). T. D. J has been appointed as an Outstanding Talent by Henan Normal University.

FundersFunder number
Henan Normal University2020ZD01
National Natural Science Foundation of China22107089, 21925802, 52271251, U21A20314
Science and Technology Department of Henan Province23IRTSTHN002

    ASJC Scopus subject areas

    • General Chemistry

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