Abstract：
Biotechnological applications of phytocystatins have garnered significant interest due to their potential applications in crop protection and improve crop resistance to abiotic stress factors. Cof1 and Wal1 are phytocystatins derived from Coffea arabica and Juglans regia, respectively. These plants hold significant economic value due to coffee\'s global demand and the walnut tree\'s production of valuable timber and widely consumed walnuts with culinary and nutritional benefits. The study involved the heterologous expression in E. coli Lemo 21(DE3), purification by immobilized metal ion affinity and size exclusion chromatography, and biophysical characterization of both phytocystatins, focusing on isolating and interconverting their monomers and dimers. The crystal structure of the domain-swapped dimer of Wal1 was determined revealing two domain-swapped dimers in the asymmetric unit, an arrangement reminiscent of the human cystatin C structure. Alphafold models of monomers and Alphafold-Multimer models of domain-swapped dimers of Cof1 and Wal1 were analyzed in the context of the crystal structure. The methodology and data presented here contribute to a deeper understanding of the oligomerization mechanisms of phytocystatins and their potential biotechnological applications in agriculture.
摘要：
植物细胞抑制素的生物技术应用由于其在作物保护和提高作物对非生物胁迫因素的抗性方面的潜在应用而引起了极大的兴趣。Cof1和Wal1是来自阿拉伯咖啡和核桃属植物细胞抑制素，分别。由于咖啡的全球需求和核桃树生产的珍贵木材和广泛食用的核桃具有烹饪和营养益处，这些植物具有重要的经济价值。该研究涉及在大肠杆菌Lemo21（DE3）中的异源表达，通过固定化金属离子亲和和尺寸排阻色谱法纯化，和两种植物细胞抑制素的生物物理表征，专注于分离和相互转化它们的单体和二聚体。确定了Wal1的结构域交换二聚体的晶体结构，揭示了不对称单元中的两个结构域交换二聚体，让人联想到人类胱抑素C结构的排列。在晶体结构的背景下分析了Cof1和Wal1的单体的α折叠模型和结构域交换二聚体的α折叠-多聚体模型。此处提供的方法和数据有助于更深入地了解植物细胞抑制素的低聚机制及其在农业中的潜在生物技术应用。
