Department of Biosciences and Nutrition, Karolinska Institutet, Neo, Blickagången 16, Huddinge, 141 52, Sweden.
Department of Physical Organic Chemistry, Latvian Institute of Organic Synthesis, Riga, LV-1006, Latvia.
Nat Commun. 2022 Aug 15;13(1):4695. doi: 10.1038/s41467-022-32093-7.
Recombinant spider silk proteins (spidroins) have multiple potential applications in development of novel biomaterials, but their multimodal and aggregation-prone nature have complicated production and straightforward applications. Here, we report that recombinant miniature spidroins, and importantly also the N-terminal domain (NT) on its own, rapidly form self-supporting and transparent hydrogels at 37 °C. The gelation is caused by NT α-helix to β-sheet conversion and formation of amyloid-like fibrils, and fusion proteins composed of NT and green fluorescent protein or purine nucleoside phosphorylase form hydrogels with intact functions of the fusion moieties. Our findings demonstrate that recombinant NT and fusion proteins give high expression yields and bestow attractive properties to hydrogels, e.g., transparency, cross-linker free gelation and straightforward immobilization of active proteins at high density.
重组蜘蛛丝蛋白（丝氨酸）在新型生物材料的开发中有多种潜在应用，但它们的多模态和易于聚集的性质使得其生产和直接应用变得复杂。在这里，我们报告说，重组微型丝氨酸，以及重要的是其自身的 N 端结构域（NT），可以在 37°C 下迅速形成自支撑且透明的水凝胶。凝胶化是由 NT α-螺旋向 β-折叠的转换以及类似淀粉样纤维的形成引起的，并且由 NT 和绿色荧光蛋白或嘌呤核苷磷酸化酶组成的融合蛋白形成具有融合部分完整功能的水凝胶。我们的研究结果表明，重组 NT 和融合蛋白具有较高的表达产量，并赋予水凝胶吸引人的特性，例如透明度、无交联剂的凝胶化以及高浓度下活性蛋白的简单固定。
