Investigating spider silk self-assembly via liquid-liquid phase separation

今年，与Kakeni赠款有关的工作取得了重大进展。与MaSp1组装相关的实验工作已经完成，包括研究蛋白质的2D结构以及在介观尺度上实时表征完整的自组装过程。这些结果已经发表在一篇论文中。不同MASP变体的复合功能条件的优化工作已经在小规模上取得了成功，但大规模的纤维纺丝和表征工作尚未成功完成。已经对具有重复结构域氨基酸基序变异的MaSp2变体阵列进行了分析，包括在Spring-8使用SEC-SAXS系统进行了数据收集和分析；此外，还进行了探索突变对LLP可逆性/效率的影响的实验，尽管这些实验尚未最后敲定。这些工作的成果目前正在撰写出版。成功发表了3篇关于蜘蛛丝结构/功能的同行评议论文(包括1篇作为第一作者之一)，并参加了会议。

This year significant progress was achieved on the work related to the Kakenhi grant. The experimental work related to the investigation on MaSp1 assembly (in isolation) has been completed, including work on the 2D protein structure as well as real-time characterization of the complete self-assembly process at the mesoscale. These results have been written up in a paper for publication.Work on the optimization of conditions for the composite function of the different MaSp variants have been carried out, which were successful on the small-scale, however the large-scale fiber spinning and characterization has not yet been successfully completed. The analysis of the array of MaSp2 variants with variations in the repetitive domain amino acid motifs have been carried, including data collection and analysis at SPring-8 using the SEC-SAXS system, have been performed; in addition, experiments exploring the effect of mutations on reversibility/efficiency of LLPS were carried out, although these have not been finalized. The results of these works are currently being written up for publication.Three peer-reviewed papers related to spider silk structure/function were successfully published (including 1 as co-first author), and conferences were attended.