A Ratiometric Two‐Photon Fluorescent Probe for Tracking Lysosomal ATP: Direct In Cellulo Observation of Lysosomal Membrane Fusion Processes,Angewandte Chemie International Edition

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A Ratiometric Two‐Photon Fluorescent Probe for Tracking Lysosomal ATP: Direct In Cellulo Observation of Lysosomal Membrane Fusion Processes
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2018-07-17 , DOI: 10.1002/anie.201804743
Yong Woong Jun ₁ , Taejun Wang ₂ , Sekyu Hwang ₁ , Dokyoung Kim _{3,

4} , Donghee Ma ₁ , Ki Hean Kim ₂ , Sungjee Kim ₁ , Junyang Jung ₃ , Kyo Han Ahn ₁

Affiliation

Vesicles exchange their contents through membrane fusion processes, kiss‐and‐run and full‐collapse fusion. Indirect observation of these fusion processes using artificial vesicles enhanced our understanding on the molecular mechanisms involved. Direct observation of the fusion processes in a real biological system, however, remains a challenge owing to many technical obstacles. We report a ratiometric two‐photon probe offering real‐time tracking of lysosomal ATP with quantitative information for the first time. By applying the probe to two‐photon live‐cell imaging, the lysosomal membrane fusion process in cells has been directly observed and the concentration of its content, lysosomal ATP, has been measured. Results show that the kiss‐and‐run process between lysosomes proceeds through repeated transient interactions with gradual content mixing, whereas the full‐fusion process occurs at once. Furthermore, it is confirmed that both the fusion processes proceed with conservation of the content. Such a small‐molecule probe exerts minimal disturbance and hence has potential for studying various biological processes associated with lysosomal ATP.

中文翻译：

用于跟踪溶酶体ATP的比例双光子荧光探针：直接在纤维素中观察溶酶体膜融合过程。

囊泡通过膜融合过程，亲吻和奔跑以及完全塌陷的融合来交换其内含物。使用人工囊泡对这些融合过程的间接观察增强了我们对涉及的分子机制的理解。然而，由于许多技术障碍，在真实生物系统中直接观察融合过程仍然是一个挑战。我们首次报道了一种比例式双光子探针，可实时跟踪溶酶体ATP并提供定量信息。通过将探针应用于双光子活细胞成像，可以直接观察到细胞中的溶酶体膜融合过程，并测量了其溶酶体ATP的浓度。结果表明，溶酶体之间的亲吻和奔跑过程是通过反复的短暂相互作用与逐渐混合的内容物进行的，而全融合过程会立即发生。此外，证实了两种融合过程均进行了内容物的保存。这种小分子探针产生的干扰极小，因此具有研究与溶酶体ATP相关的各种生物学过程的潜力。

更新日期：2018-07-17

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