Wang Yafu, Niu Huiyu, Wang Kui, Yang Liu, Wang Ge, James Tony D, Fan Jiangli, Zhang Hua
Collaborative Innovation Centre of Henan Province for Green Manufacturing of Fine Chemicals, Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, Henan Key Laboratory of Organic Functional Molecule and Drug Innovation, School of Chemistry and Chemical Engineering, Henan Normal University Xinxiang Henan 453007 P. R. China
Xinxiang Medical University Xinxiang 453000 P. R. China.
Chem Sci. 2024 Sep 12;15(39):16291-9. doi: 10.1039/d4sc03387f.
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 AC-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.
转移酶是具有多位点迁移特性的酶。重要的是，酶活性在这个迁移过程中会发生变化，这不可避免地会影响生物体的生理功能，甚至可能导致相关的恶性疾病。然而，该领域的研究因缺乏同时、差异监测位点特异性转移酶活性的工具而受到严重阻碍。在此，我们提出了一种新策略，该策略整合了具有高灵敏度的荧光信号响应和具有卓越空间分辨率的旋光信号响应。为了验证该策略的可行性，以转移酶γ-谷氨酰转移酶（GGT）为模型系统，使用手性氨基酸作为特定的双功能识别基团，开发了双模式手性探针AC-GGTB（AC17-GGTB和AC15-GGTB）。探针在GGT作用下发生结构变化，导致双功能识别基团（手性氨基酸）释放，同时产生荧光信号和旋光信号。这种双模式输出具有高灵敏度，便于位点区分。此外，它能够在迁移过程中同时、差异检测不同位点的GGT活性。我们预计基于该策略开发的探针将有助于基于成像监测其他转移酶的活性，从而提供一个适合实时跟踪迁移过程中转移酶活性变化的成像平台。
