Labeling Lysosomes and Tracking Lysosome-Dependent Apoptosis with a Cell-Permeable Activity-Based Probe

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• 作者：Fengkai Fan ; Si Nie ; Dongmei Yang ; Meijie Luo ; Hua Shi ; Yu-Hui Zhang
• 刊名：Bioconjugate Chemistry
• 出版年：2012
• 出版时间：June 20, 2012
• 年：2012
• 卷：23
• 期：6
• 页码：1309-1317
• 全文大小：436K
• 年卷期：v.23,no.6(June 20, 2012)
• ISSN：1520-4812

文摘

In this study, we describe a new strategy for labeling and tracking lysosomes with a cell-permeable fluorescent activity-based probe (CpFABP) that is covalently bound to select lysosomal proteins. Colocalization studies that utilized LysoTracker probes as standard lysosomal markers demonstrated that our novel probe is effective in specifically labeling lysosomes in various kinds of live cells. Furthermore, our studies revealed that this probe has the ability to label fixed cells, permeabilized cells, and NH₄Cl-treated cells, unlike LysoTracker probes, which show ineffective labeling under the same conditions. Remarkably, when applied to monitor the process of lysosome-dependent apoptosis, our probe not only displayed the expected release of lysosomal cathepsins from lysosomes into the cytosol but also revealed additional information about the location of the cathepsins during apoptosis, which is undetectable by other chemical lysosome markers. These results suggest a wide array of promising applications for our probe and provide useful guidelines for its use as a lysosome marker in lysosome-related studies.