Single-molecule displacement mapping unveils nanoscale heterogeneities in intracellular diffusivity.,Nature Methods

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Single-molecule displacement mapping unveils nanoscale heterogeneities in intracellular diffusivity.
Nature Methods ( IF 36.1 ) Pub Date : 2020-03-16 , DOI: 10.1038/s41592-020-0793-0
Limin Xiang _{1,

2} , Kun Chen _{1,

2} , Rui Yan _{1,

2} , Wan Li _{1,

2} , Ke Xu _{1,

2}

Affiliation

Intracellular diffusion underlies vital cellular processes. However, it remains difficult to elucidate how an unbound protein diffuses inside the cell with good spatial resolution and sensitivity. Here we introduce single-molecule displacement/diffusivity mapping (SMdM), a super-resolution strategy that enables the nanoscale mapping of intracellular diffusivity through local statistics of the instantaneous displacements of freely diffusing single molecules. We thus show that the diffusion of an average-sized protein in the mammalian cytoplasm and nucleus is spatially heterogeneous at the nanoscale, and that variations in local diffusivity correlate with the ultrastructure of the actin cytoskeleton and the organization of the genome, respectively. SMdM of differently charged proteins further unveils that the possession of positive, but not negative, net charges drastically impedes diffusion, and that the rate is determined by the specific subcellular environments. We thus unveil rich heterogeneities and charge effects in intracellular diffusion at the nanoscale.

中文翻译：

单分子置换图谱揭示了细胞内扩散率的纳米级异质性。

细胞内扩散是重要的细胞过程的基础。然而，仍然难以阐明未结合的蛋白质如何以良好的空间分辨率和灵敏度在细胞内扩散。在这里，我们介绍单分子位移/扩散图谱（SMdM），这是一种超分辨率策略，通过对自由扩散的单个分子的瞬时位移进行局部统计，可以对细胞内扩散率进行纳米级映射。因此，我们表明，平均大小的蛋白质在哺乳动物细胞质和细胞核中的扩散在纳米尺度上是空间异质的，并且局部扩散率的变化分别与肌动蛋白细胞骨架的超微结构和基因组的组织有关。带有不同电荷的蛋白质的SMdM进一步揭示了拥有阳性但不是阴性的蛋白质净电荷极大地阻止了扩散，并且速率由特定的亚细胞环境决定。因此，我们揭示了纳米级细胞内扩散中的丰富异质性和电荷效应。

更新日期：2020-03-16

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