A Method to Quantify Molecular Diffusion within Thin Solvated Polymer Films: A Case Study on Films of Natively Unfolded Nucleoporins.
A Method to Quantify Molecular Diffusion within Thin Solvated Polymer Films: A Case Study on Films of Natively Unfolded Nucleoporins.
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DOI：
10.1021/acsnano.0c02895
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发表时间：
2020-08-25
期刊：
影响因子：
17.1
通讯作者：
Richter RP
中科院分区：
文献类型：
--
作者：
Frost R;Débarre D;Jana S;Bano F;Schünemann J;Görlich D;Richter RP
我们提出了一种方法来探测分子和纳米粒子扩散薄，溶剂化聚合物涂层。该装置利用了在紧密接触的平面和半球形表面（其中至少一个表面涂覆有聚合物）之间的界面处形成的具有良好限定的几何形状的限制，并使用该限制来分析扩散过程，而不干扰与本体溶液的交换和扩散。用这种方法，我们称之为平面球限制显微镜（PSCM），关于聚合物涂层和散装液体之间的分子的分区的信息也得到了。由于受限的几何形状，扩散和分配可以在单个实验中映射为涂层的压缩和浓度的函数。该方法是通用的，可以与传统的光学显微镜集成，因此，它应该发现在许多应用领域利用功能聚合物涂层的广泛使用。我们证明了使用刷的天然未折叠的核孔蛋白结构域丰富的苯丙氨酸-甘氨酸重复序列（FG域）的PSCM。已知FG结构域的网状结构负责核转运受体（NTR）及其大分子货物穿过分离活细胞的胞质溶胶和细胞核的核膜的选择性转运。我们发现，选择性的NTR吸收FG域膜依赖于FG域浓度敏感，NTRs与FG域的相互作用阻碍NTR运动只有适度。这些观察有助于重要的信息，以更好地了解选择性NTR运输的机制。
We present a method to probe molecular and nanoparticle diffusion within thin, solvated polymer coatings. The device exploits the confinement with well-defined geometry that forms at the interface between a planar and a hemispherical surface (of which at least one is coated with polymers) in close contact and uses this confinement to analyze diffusion processes without interference of exchange with and diffusion in the bulk solution. With this method, which we call plane–sphere confinement microscopy (PSCM), information regarding the partitioning of molecules between the polymer coating and the bulk liquid is also obtained. Thanks to the shape of the confined geometry, diffusion and partitioning can be mapped as a function of compression and concentration of the coating in a single experiment. The method is versatile and can be integrated with conventional optical microscopes; thus it should find widespread use in the many application areas exploiting functional polymer coatings. We demonstrate the use of PSCM using brushes of natively unfolded nucleoporin domains rich in phenylalanine–glycine repeats (FG domains). A meshwork of FG domains is known to be responsible for the selective transport of nuclear transport receptors (NTRs) and their macromolecular cargos across the nuclear envelope that separates the cytosol and the nucleus of living cells. We find that the selectivity of NTR uptake by FG domain films depends sensitively on FG domain concentration and that the interaction of NTRs with FG domains obstructs NTR movement only moderately. These observations contribute important information to better understand the mechanisms of selective NTR transport.
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影响因子：
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作者：
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