Zhang Xiao-Fan, Wang Tian-Ran, Cao Xiao-Qun, Shen Shi-Li
Taian Center For Food and Drug Control, Taian 271000, PR China.
School of Chemistry and Pharmaceutical Engineering, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian 271016, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2020 Feb 15;227:117761. doi: 10.1016/j.saa.2019.117761. Epub 2019 Nov 6.
Heat shock is a potentially fatal condition characterized by high body temperature (>40 °C), which may lead to physical discomfort and dysfunctions of organ systems. Acidic pH environment in lysosomes can activate enzymes, thus facilitating the degradation of proteins in cellular metabolism. Owing to the lack of a practical research tool, it remains difficult to exploit relationship between heat shock and lysosome. Herein, a NIR lysosomal pH chemosensor (NRLH) was developed. One typical lysosome-locating group, morpholine, was incorporated into NRLH. The fluorescence intensity showed pH-dependent characteristics and responded sensitively to pH fluctuations in the pH range of 3.0-5.5. NRLH with a pKa of 4.24 displayed rapid response and high selectivity for H among common species. We also demonstrated NRLH was capable of targeting lysosomes. Importantly, NRLH was applied in cellular imaging and the data revealed that lysosomal pH increased but never decreased during the heat shock. Therefore, NRLH may act as an effective molecular tool for exploring the mechanisms of heat-related pathology in bio-systems.
热休克是一种潜在的致命状况,其特征为体温过高(>40°C),这可能会导致身体不适以及器官系统功能障碍。溶酶体中的酸性pH环境可激活酶,从而促进细胞代谢中蛋白质的降解。由于缺乏实用的研究工具,探索热休克与溶酶体之间的关系仍然困难重重。在此,我们开发了一种近红外溶酶体pH化学传感器(NRLH)。一个典型的溶酶体定位基团吗啉被引入到NRLH中。荧光强度呈现出pH依赖性特征,并对3.0 - 5.5 pH范围内的pH波动做出灵敏响应。pKa为4.24的NRLH对常见物种中的H+显示出快速响应和高选择性。我们还证明了NRLH能够靶向溶酶体。重要的是,NRLH被应用于细胞成像,数据显示在热休克期间溶酶体pH升高但从未降低。因此,NRLH可能作为一种有效的分子工具来探索生物系统中热相关病理学的机制。