Data from: Peroxiredoxin is a Versatile Self-Assembling Tecton for Protein Nanotechnology

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The potential for protein tectons to be used in nanotechnology is increasingly recognized, but the repertoire of stable proteins that assemble into defined shapes in response to an environmental trigger is limited. Peroxiredoxins (Prxs) are a protein family that shows an amazing array of supramolecular assemblies, making them attractive tectons. Human Prx3 (hPrx3) forms toroidal oligomers characteristic of the Prx family, but no structure has been solved to date. Here we report the first 3-D structure of this protein, derived from single-particle analysis of TEM images, establishing a dodecameric structure. This result was supported by SAXS measurements. We also present the first detailed structure of a double toroidal Prx from a higher organism determined by SPA. Guided by these structures, variants of the protein were designed to facilitate controlled assembly of protein nanostructures through the association of the toroids. We observed an enhanced population of stacked toroids, as seen by TEM; nanocages and interlocked toroids were also visible. Low pH was successfully predicted to generate long ordered nanotubes. Control over the length of the tubes was gained by adding ammonium sulfate to the assembly buffer. These versatile assembly properties demonstrate the considerable potential of hPrx3 as a tecton for protein nanotechnology.

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蛋白质结构单元（protein tectons）在纳米技术中的应用潜力日益得到学界认可，但能够响应环境触发因素、组装为明确构型的稳定蛋白质库仍十分有限。过氧化物氧还蛋白（Peroxiredoxins, Prxs）是一类蛋白质家族，展现出丰富多样的超分子组装形式，因此成为极具吸引力的结构单元。人类Prx3（hPrx3）可形成Prx家族特有的环形寡聚体，但迄今为止尚未解析其三维结构。本研究首次解析了该蛋白质的三维结构：通过透射电子显微镜（Transmission Electron Microscopy, TEM）图像的单颗粒分析，确定其为十二聚体结构，该结果得到了小角X射线散射（Small Angle X-ray Scattering, SAXS）实验的验证。本研究还报道了首个来自高等生物的双环形Prx的精细结构，该结构通过单颗粒分析（Single-Particle Analysis, SPA）解析得到。

基于上述结构，研究人员设计了该蛋白质的突变体，以通过环形亚基的相互结合实现蛋白质纳米结构的可控组装。透射电子显微镜观测显示，堆叠环形寡聚体的数量显著增多，同时还观察到了纳米笼与互锁环形结构。研究成功预测低pH环境可诱导形成长程有序的纳米管，并通过在组装缓冲液中添加硫酸铵实现了对纳米管长度的精准调控。这些多样的组装特性证明了hPrx3作为蛋白质纳米技术结构单元的巨大应用潜力。