Labeling Lysosomes and Tracking Lysosome-Dependent Apoptosis with a Cell-Permeable Activity-Based Probe
Labeling Lysosomes and Tracking Lysosome-Dependent Apoptosis with a Cell-Permeable Activity-Based Probe
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DOI：
10.1021/bc300143p
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发表时间：
2012-06-01
影响因子：
4.7
通讯作者：
Zhang, Yu-Hui
中科院分区：
文献类型：
--
作者：
Fan, Fengkai;Nie, Si;Zhang, Yu-Hui
在这项研究中，我们描述了一种使用细胞可渗透的基于荧光活性的探针（CpFABP）标记和跟踪溶酶体的新策略，该探针与选定的溶酶体蛋白共价结合。使用 LysoTracker 探针作为标准溶酶体标记物的共定位研究表明，我们的新型探针可有效特异性标记各种活细胞中的溶酶体。此外，我们的研究表明，该探针能够标记固定细胞、透化细胞和 NH4Cl 处理的细胞，这与 LysoTracker 探针不同，后者在相同条件下显示出无效标记。值得注意的是，当应用于监测溶酶体依赖性细胞凋亡过程时，我们的探针不仅显示了溶酶体组织蛋白酶从溶酶体释放到细胞质中的预期，而且还揭示了有关细胞凋亡过程中组织蛋白酶位置的其他信息，这是其他化学溶酶体标记物无法检测到的。这些结果表明我们的探针具有广泛的有前途的应用，并为其在溶酶体相关研究中用作溶酶体标记物提供了有用的指导。
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.