Nanoscale Visualization of Morphological Alteration of Live-Cell Membranes by the Interaction with Oligoarginine Cell-Penetrating Peptides
Nanoscale Visualization of Morphological Alteration of Live-Cell Membranes by the Interaction with Oligoarginine Cell-Penetrating Peptides
复制标题
DOI：
10.1021/acs.analchem.0c04097
复制
发表时间：
2021-03-26
影响因子：
7.4
通讯作者：
Matsue, Tomokazu
中科院分区：
文献类型：
--
作者：
Ida, Hiroki;Takahashi, Yasufumi;Matsue, Tomokazu
细胞膜和生物分子之间的相互作用仍然知之甚少。例如，富含精氨酸的细胞穿透肽(CPPs)，包括八精氨酸(R8)，通过与细胞膜的相互作用而内化。然而，在内化过程中，这些CPP引入的确切的膜动力学仍不清楚。在这里，我们使用一个结合了扫描离子电导显微镜和旋转圆盘共聚焦显微镜的系统，使用荧光标记的R8来可视化富含精氨酸的CPP和细胞-膜相互作用引起的形态变化。该系统允许对活细胞膜中的结构动力学进行随时间变化的纳米级可视化。因此，可以观察到富含精氨酸的CPPS引起的各种类型的膜重构。膜褶皱的诱导和杯状闭合被观察到是一个内吞多肽的过程。另一种建议是凹形结构形成，伴随着多肽通过细胞膜的直接转位。在没有荧光标记的情况下使用R8的研究也证明了荧光部分对膜结构改变的不可忽视的影响。
The interactions between the cell membrane and biomolecules remain poorly understood. For example, arginine-rich cell-penetrating peptides (CPPs), including octaarginines (R8), are internalized by interactions with cell membranes. However, during the internalization process, the exact membrane dynamics introduced by these CPPs are still unknown. Here, we visualize arginine-rich CPPs and cell-membrane interaction-induced morphological changes using a system that combines scanning ion-conductance microscopy and spinning-disk confocal microscopy, using fluorescently labeled R8. This system allows time-dependent, nanoscale visualization of structural dynamics in live-cell membranes. Various types of membrane remodeling caused by arginine-rich CPPs are thus observed. The induction of membrane ruffling and the cup closure are observed as a process of endocytic uptake of the peptide. Alternatively suggested is the concave structural formation accompanied by direct peptide translocation through cell membranes. Studies using R8 without fluorescent labeling also demonstrate a non-negligible effect of the fluorescent moiety on membrane structural alteration.