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

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Fluorescence-plane polarization for the real-time monitoring of transferase migration in living cells
Chemical Science ( IF 7.6 ) Pub Date : 2024-09-12 , DOI: 10.1039/d4sc03387f
Yafu Wang ₁ , Huiyu Niu ₁ , Kui Wang ₁ , Liu Yang ₁ , Ge Wang ₂ , Tony D. James _{1,

3} , Jiangli Fan ₄ , Hua Zhang ₁

Affiliation

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.

中文翻译：

荧光平面极化，用于实时监测活细胞中转移酶的迁移

转移酶是表现出多位点迁移特性的酶。值得注意的是，酶活性在这个迁移过程中会发生变化，这不可避免地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，由于缺乏同时和差异监测位点特异性转移酶活性的工具，该领域的研究受到严重阻碍。在此，我们提出了一种新的策略，该策略集成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，使用转移酶 γ-谷氨酰转移酶（GGT）作为模型系统，使用手性氨基酸作为特异性双功能识别组来开发双模式手性探针 AC x-GGTB （AC17-GGTB 和 AC15-GGTB）。探针在 GGT 下发生结构变化，导致双功能识别基团（手性氨基酸）的释放，同时产生荧光信号和旋光信号。这种双模式输出表现出高灵敏度，有助于位点的区分。此外，它能够在迁移过程中同时和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对其他转移酶的活性进行基于成像的监测，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。

更新日期：2024-09-12

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