Microscale Diffusiophoresis of Proteins.
Microscale Diffusiophoresis of Proteins.
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DOI：
10.1021/acs.jpcb.2c04029
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发表时间：
2022-11-10
期刊：
影响因子：
--
通讯作者：
Knowles TPJ
中科院分区：
文献类型：
--
作者：
Peter QAE;Jacquat RPB;Herling TW;Challa PK;Kartanas T;Knowles TPJ
生命系统的特点是其空间高度不均匀的性质，容易从根本上改变生物分子物种的行为，包括支撑细胞生物功能的蛋白质。众所周知，化学势的空间梯度会导致胶体颗粒的强运输效应，但它们对分子尺度物种（如蛋白质）的影响在很大程度上仍未被探索。在此，我们改进了现有的扩散电泳微流体技术，在真实空间中测量蛋白质扩散电泳。通过两项改进，蛋白质的测量成为可能。首先，使用无标签显微镜来抑制标签干扰。其次，数值方法的改进是为了满足小分子所带来的特殊挑战。我们证明了单个蛋白质可以在盐梯度中以一种足以克服扩散的方式进行强烈的扩散泳运动，并导致其在细胞尺度上的空间组织发生巨大变化。此外，我们证明了这种现象可以用于控制微流体装置中蛋白质的运动。这些结果为对生物系统中梯度在大分子空间组织中的作用的物理理解开辟了一条道路，并突出了在设备上实现蛋白质分选应用的新途径。
Living systems are characterized by their spatially highly inhomogeneous nature which is susceptible to modify fundamentally the behavior of biomolecular species, including the proteins that underpin biological functionality in cells. Spatial gradients in chemical potential are known to lead to strong transport effects for colloidal particles, but their effect on molecular scale species such as proteins has remained largely unexplored. Here, we improve on existing diffusiophoresis microfluidic technique to measure protein diffusiophoresis in real space. The measurement of proteins is made possible by two ameliorations. First, a label-free microscope is used to suppress label interference. Second, improvements in numerical methods are developed to meet the particular challenges posed by small molecules. We demonstrate that individual proteins can undergo strong diffusiophoretic motion in salt gradients in a manner which is sufficient to overcome diffusion and which leads to dramatic changes in their spatial organization on the scale of a cell. Moreover, we demonstrate that this phenomenon can be used to control the motion of proteins in microfluidic devices. These results open up a path towards a physical understanding of the role of gradients in living systems in the spatial organization of macromolecules and highlight novel routes towards protein sorting applications on device.
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