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微流控在研究肌动蛋白生物化学和生物力学方面的优势。

The advantages of microfluidics to study actin biochemistry and biomechanics.

机构信息

Institut Jacques Monod, CNRS, Université de Paris, 75013, Paris, France.

出版信息

J Muscle Res Cell Motil. 2020 Mar;41(1):175-188. doi: 10.1007/s10974-019-09564-4. Epub 2019 Nov 20.

Abstract

The regulated assembly of actin filaments is essential in nearly all cell types. Studying actin assembly dynamics can pose many technical challenges. A number of these challenges can be overcome by using microfluidics to observe and manipulate single actin filaments under an optical microscope. In particular, microfluidics can be tremendously useful for applying different mechanical stresses to actin filaments and determining how the physical context of the filaments affects their regulation by biochemical factors. In this review, we summarize the main features of microfluidics for the study of actin assembly dynamics, and we highlight some recent developments that have emerged from the combination of microfluidics and other techniques. We use two case studies to illustrate our points: the rapid assembly of actin filaments by formins and the disassembly of filaments by actin depolymerizing factor (ADF)/cofilin. Both of these protein families play important roles in cells. They regulate actin assembly through complex molecular mechanisms that are sensitive to the filaments' mechanical context, with multiple activities that need to be quantified separately. Microfluidics-based experiments have been extremely useful for gaining insight into the regulatory actions of these two protein families.

摘要

肌动蛋白丝的调控组装对于几乎所有细胞类型都是至关重要的。研究肌动蛋白组装动力学可能会带来许多技术挑战。通过使用微流控技术在光学显微镜下观察和操纵单个肌动蛋白丝,可以克服其中的许多挑战。特别是,微流控技术在向肌动蛋白丝施加不同机械应力并确定丝的物理环境如何影响其被生化因子调节方面非常有用。在这篇综述中,我们总结了微流控技术在研究肌动蛋白组装动力学方面的主要特点,并强调了一些最新的发展,这些发展是通过微流控技术与其他技术的结合而产生的。我们使用两个案例研究来说明我们的观点:formin 家族快速组装肌动蛋白丝和肌动蛋白解聚因子 (ADF)/丝切蛋白解聚肌动蛋白丝。这两个蛋白家族在细胞中都起着重要作用。它们通过对丝的机械环境敏感的复杂分子机制来调节肌动蛋白的组装,其中许多活性需要单独进行量化。基于微流控的实验对于深入了解这两个蛋白家族的调节作用非常有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/283a/7109186/c49dea61fca8/10974_2019_9564_Fig1_HTML.jpg

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