Long Zhengze, Li Yang, Yu Ting, Li Ting, Wang Jianhua, Zhang Youyu, Li Haitao, Yin Peng, Zhou Huijun, Yao Shouzhuo
Department of Gastroenterology and Urology, The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University/Hunan Cancer Hospital, Changsha 410013, P.R. China.
Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education), Institute of Interdisciplinary Studies, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, China.
Anal Chem. 2025 Jul 15;97(27):14620-14628. doi: 10.1021/acs.analchem.5c02130. Epub 2025 Jul 2.
Lysosomal cysteine (Cys), glutathione (GSH), and homocysteine (Hcy) are vital biothiols essential for maintaining cellular redox homeostasis and regulating critical physiological functions. Aberrant levels of these biothiols are closely linked to various diseases. Understanding the dynamics of lysosomal biothiols is crucial for elucidating their physiological roles and involvement in disease progression. However, the simultaneous and selective detection of Cys, GSH, and Hcy in lysosomes remains challenging due to their structural similarities and dynamic behaviors. In this study, we developed , a novel coumarin-based fluorescent probe tailored for lysosomal targeting and the simultaneous discrimination of Cys, GSH, and Hcy through three distinct emission channels. By integrating multiple binding sites with a lysosome-targeting moiety, exhibits high specificity, sensitivity, and subcellular selectivity, enabling precise visualization of biothiol dynamics under oxidative and hyperosmotic stress conditions. Additionally, was successfully employed for real-time monitoring of biothiol fluctuations in a zebrafish model of oxidative and hyperosmotic stress. This work introduces an innovative tool for studying lysosomal biothiol dynamics, offering critical insights into lysosome-associated processes and their roles in cellular physiology and pathology. Moreover, it presents a promising strategy for diagnosing and investigating lysosome-associated diseases, paving the way for future biomedical and therapeutic advancements.
溶酶体中的半胱氨酸（Cys）、谷胱甘肽（GSH）和同型半胱氨酸（Hcy）是维持细胞氧化还原稳态和调节关键生理功能所必需的重要生物硫醇。这些生物硫醇水平异常与多种疾病密切相关。了解溶酶体生物硫醇的动态变化对于阐明其生理作用以及在疾病进展中的参与情况至关重要。然而，由于它们的结构相似性和动态行为，同时选择性检测溶酶体中的Cys、GSH和Hcy仍然具有挑战性。在本研究中，我们开发了一种新型的基于香豆素的荧光探针，该探针专为溶酶体靶向设计，并通过三个不同的发射通道同时区分Cys、GSH和Hcy。通过将多个结合位点与溶酶体靶向部分整合，该探针具有高特异性、灵敏度和亚细胞选择性，能够在氧化和高渗应激条件下精确可视化生物硫醇的动态变化。此外，该探针成功用于在氧化和高渗应激斑马鱼模型中实时监测生物硫醇的波动。这项工作引入了一种研究溶酶体生物硫醇动态变化的创新工具，为深入了解溶酶体相关过程及其在细胞生理和病理中的作用提供了关键见解。此外，它还为诊断和研究溶酶体相关疾病提供了一种有前景的策略，为未来的生物医学和治疗进展铺平了道路。
