Self-assembly und structure formation of spider silk proteins in (ultra-)thin films

（超）薄膜中蜘蛛丝蛋白的自组装和结构形成

• 批准号: 410872515
• 负责人: Professor Dr. Andreas Fery
• 金额: --
• 依托单位: Leibniz-Institut für Polymerforschung Dresden (IPF)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/410872515?language=en
• 关键词: Self; assembly; und; structure; formation

The preparation of (ultra) thin spider silk films (d = 2 – 100 nm) in combination with the design of various (new) recombinant spider silk proteins offers the unique possibility to investigate the structure formation and self-assembly behavior of silk proteins in thin films at defined interfaces and allows to compare these processes to those in bulk solution. The coating of the substrates and preparation of the films is achieved by deposition from aqueous solution. By self-assembly as well as surface interaction phase separated structures with domains in the nanoscale are formed. This proposal aims at the elucidation of structure formation within spider silk proteins in dependence of their amino acid sequence (amino acid composition as well as sequence of defined amino acid modules). The general question of this proposal addresses the modalities of deposition and orientation of spider silk-related films as well as the influence of primary structure (i.e. sequence), of amino acid charge, and of the molecular weight of spider silk proteins in (ultra-)thin films, as well as the influence of surface chemistry and topography of the used support materials. The size of the gained structural nano-sized domains will be controlled via the sequence of the modules. The following aims are tackled in detail:• Time course (kinetics, dynamics) of the folding (secondary structure) and orientation of the structural elements in spider silk protein films at surfaces• Influence of charge (exchange of amino acid E by K, from C- to Kappa-module) and its relation to salinity and pH• Role of the typically major fraction of the random coil/alpha-helix rich hydrophilic amorphous phase concerning the formation of the minor fraction of beta-sheet rich hydrophobic crystalline phases within spider silk materials (embedding) • Role of the aqueous (buffer, pH, salinity) respectively organic solvent• Role of mechanical textures, orientation and surface properties of substrates concerning structure formation and orientation of the domainsThe characterization of the orientation of secondary structural elements by polarised FTIR spectroscopy and of the separation of spider silk peptide blocks by high resolution atomic force microscopy techniques allows to draw conclusions with respect to the mechanism of structure formation and to create a general assembly model of spider silk based thin films.

（超）薄蜘蛛丝薄膜（d = 2 - 100 nm）的制备与各种（新的）重组蜘蛛丝蛋白的设计相结合，为研究薄膜中丝蛋白在定义界面处的结构形成和自组装行为提供了独特的可能性，并允许将这些过程与体溶液中的过程进行比较。基材的涂层和薄膜的制备是通过从水溶液中沉积来实现的。通过自组装和表面相互作用，形成了具有纳米级畴的相分离结构。本提案旨在阐明蜘蛛丝蛋白的结构形成依赖于其氨基酸序列（氨基酸组成以及定义的氨基酸模块的序列）。本提案的一般问题涉及蜘蛛丝相关薄膜的沉积和取向方式，以及（超）薄膜中蜘蛛丝蛋白的一级结构（即序列）、氨基酸电荷和分子量的影响，以及所用支撑材料的表面化学和形貌的影响。获得的纳米结构域的大小将通过模块的顺序来控制。详细讨论了以下目标：•蜘蛛丝蛋白膜表面折叠（二级结构）和结构元素取向的时间过程（动力学、动力学）•电荷的影响（氨基酸E与K的交换）；从C-到kappa -模块)及其与盐度和pH的关系•蜘蛛丝材料中典型的主要部分的无规线圈/富含α -螺旋的亲水无定形相对少量富含β -薄片的疏水结晶相形成的作用（包埋）•水（缓冲液、pH值、盐度）有机溶剂的作用•机械织构的作用利用极化FTIR光谱对二级结构元素的取向进行表征，并利用高分辨率原子力显微镜技术对蜘蛛丝肽块的分离进行表征，从而得出有关结构形成机制的结论，并建立了蜘蛛丝基薄膜的总体组装模型。