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Control of stability of polypeptide multilayer nanofilms by quantitative control of disulfide bond formation

机译：通过定量控制二硫键形成来控制多肽多层纳米膜的稳定性

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The crosslinking of polymers in a polymeric material will alter the mechanical properties of the material. Control over the mechanical properties of polyelectrolyte multilayer films (PEMs) could be useful for applications of the technology in medicine and other areas. Bisulfide bonds are 'natural' polypeptide crosslinks found widely in wild-type proteins. Here, we have designed and synthesized three pairs of oppositely charged 32mer polypeptide to have 0, 4, or 8 cysteine (Cys) residues per molecule, and we have characterized physical properties of the peptides in a PEM context. The average linear density of free thiol in the designed peptides was 0, 0.125, or 0.25 per amino acid residue. The peptides were used to make 10-bilayer PEMs by electrostatic layer-by-layer self-assembly (LBL). Cys was included in the peptides to study specific effects of disulfide bond formation on PEM properties. Features of film assembly have been found to depend on the amino acid sequence, as in protein folding. Following polypeptide self-assembly into multilayer films, Cys residues were disulfide-crosslinked under mild oxidizing conditions. The stability of the crosslinked films at acidic pH has been found to depend on the number of Cys residues per peptide for a given crosslinking procedure. Crosslinked and non-crosslinked films have been analysed by ultraviolet spectroscopy (UVS), ellipsometry, and atomic force microscopy (AFM) to characterize film assembly, surface morphology, and disassembly. A selective etching model of the disassembly process at acidic pH is proposed on the basis of the experimental data. In this model, regions of film in which the disulfide bond density is low are etched at a higher rate than regions where the density is high.
机译：聚合物材料中聚合物的交联会改变材料的机械性能。控制聚电解质多层膜（PEM）的机械性能对于将该技术应用于医学和其他领域可能是有用的。双硫键是在野生型蛋白质中广泛发现的“天然”多肽交联键。在这里，我们设计并合成了三对带相反电荷的32mer多肽，每个分子具有0、4或8个半胱氨酸（Cys）残基，并且在PEM环境中表征了这些肽的物理性质。设计的肽中游离硫醇的平均线性密度为每个氨基酸残基0、0.125或0.25。该肽用于通过静电逐层自组装（LBL）制备10双层PEM。肽中包含Cys，以研究二硫键形成对PEM性能的特定影响。已经发现膜组装的特征取决于氨基酸序列，如在蛋白质折叠中。多肽自组装成多层膜后，在温和的氧化条件下将Cys残基二硫键交联。已经发现，在给定的交联过程中，交联膜在酸性pH下的稳定性取决于每个肽的Cys残基数。交联和非交联的膜已通过紫外光谱（UVS），椭圆偏振法和原子力显微镜（AFM）进行了分析，以表征膜的组装，表面形态和分解。根据实验数据，提出了在酸性pH条件下拆卸过程的选择性刻蚀模型。在该模型中，二硫键密度低的膜区域的蚀刻速度比密度高的区域高。

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