Dynamic Detection of Active Enzyme Instructed Supramolecular Assemblies In Situ via Super-Resolution Microscopy
Dynamic Detection of Active Enzyme Instructed Supramolecular Assemblies In Situ via Super-Resolution Microscopy
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通过超分辨率显微镜原位动态检测活性酶指导的超分子组装体
DOI：
10.1021/acsnano.0c00883
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发表时间：
2020
期刊：
影响因子：
17.1
通讯作者：
Gao Yuan
中科院分区：
文献类型：
--
作者：
Yao Qingxin;Wang Chenlei;Fu Meifang;Dai Luru;Li Junbai;Gao Yuan
受自然界自组装现象的启发，外源小分子在生物环境中的指令自组装已成为控制细胞命运的普遍过程。尽管越来越多的例子，多功能的生物活性，潜在的机制仍然不太清楚，这是在很大程度上阻碍了在现场识别这些动态的生物活性的困难。在这里，与直接随机光学重建显微镜，我们能够阐明的动态形态转化的酶指示的超分子生物素原位癌细胞内的分辨率低于50 nm。这表明，不同磷酸酶活性和分布的细胞系之间的组装分子经历了截然不同的途径。在HeLa细胞中，直接形成的细胞内超分子纳米纤维显示出轻微的细胞毒性，这是由于可能的细胞分泌途径，以排出这些外源分子组装体。相比之下，在Saos-2细胞表面具有活性磷酸酶的细胞中，首先在细胞膜上形成具有颗粒形态的组装体，然后转化为纳米纤维并在细胞中积聚，最终诱导Saos-2细胞死亡。总之，我们提供了一种方便的方法来揭示超分子组装体在生物环境中的原位动态纳米形态转变，以破译其多样的生物活性。
Inspired by the self-assembly phenomena in nature, the instructed self-assembly of exogenous small molecules in a biological environment has become a prevalent process to control cell fate. Despite mounting examples of versatile bioactivities, the underlying mechanism remains less understood, which is in large hindered by the difficulties in the identification of those dynamic assembliesin situ. Here, with direct stochastic optical reconstruction microscopy, we are able to elucidate the dynamic morphology transformation of the enzyme-instructed supramolecular assembliesin situinside cancer cells with a resolution below 50 nm. It indicates that the assembling molecules endure drastically different pathways between cell lines with different phosphatase activities and distribution. In HeLa cells, the direct formation of intracellular supramolecular nanofibers showed slight cytotoxicity, which was due to the possible cellular secretory pathway to excrete those exogenous molecules assemblies. In contrast, in Saos-2 cells with active phosphatase on the cell surface, assemblies with granular morphology first formed on the cell membranes, followed by a transformation into nanofibers and accumulation in cells, which induced Saos-2 cell death eventually. Overall, we provided a convenient method to reveal thein situdynamic nanomorphology transformation of the supramolecular assemblies in a biological environment, in order to decipher their diverse biological activities.