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

通过液-液相分离研究蜘蛛丝自组装

• 批准号: 21K06043
• 负责人: Malay Ali
• 金额: $ 2.66万
• 依托单位: Tokyo Institute of Technology (2022)Institute of Physical and Chemical Research (2021)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K06043/
• 关键词: silk; biomimetic; liquid phase separation; coacervate; nanofibril; self-assembly; biopolymer; biomolecular condensate; biomimetics; phase separation; spider; biomaterial; biochemistry; silk protein

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.

今年，与Kakenhi赠款有关的工作取得了重大进展。与MaSp 1组装（隔离）研究相关的实验工作已经完成，包括2D蛋白质结构以及在中尺度上完整自组装过程的实时表征。这些结果已被写在一篇论文中供发表。不同MaSp变体的复合功能的条件优化工作已经进行，这在小规模上是成功的，但是大规模的纤维纺丝和表征尚未成功完成。已对重复结构域氨基酸基序发生变化的MaSp 2变体阵列进行了分析，包括使用SEC-SAXS系统在SPring-8进行数据收集和分析;此外，还进行了探索突变对LLPS可逆性/效率影响的实验，但尚未最终确定。这些工作的成果目前正在编写出版物，成功出版了3篇关于蜘蛛丝结构/功能的同行评议论文（其中1篇为共同第一作者），并参加了会议。

项目成果

Dissecting the molecular basis for liquid-liquid phase separation of spider silk proteins during self-assembly

剖析蜘蛛丝蛋白自组装过程中液-液相分离的分子基础

• 发表时间: 2022
• 作者: Malay;Ali D.
• 通讯作者: Ali D.

Multicomponent nature underlies the extraordinary mechanical properties of spider dragline silk

• DOI: 10.1073/pnas.2107065118
• 发表时间: 2021-08-03
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Kono, Nobuaki;Nakamura, Hiroyuki;Arakawa, Kazuharu
• 通讯作者: Arakawa, Kazuharu

Exploring the self-assembly of MaSp1 dragline silk spidroin into native-like hierarchical fibers

探索 MaSp1 拖丝蜘蛛蛋白自组装成类似天然的分层纤维

• 发表时间: 2022
• 作者: Malay;Ali D.
• 通讯作者: Ali D.

Micrometer‐Scale Optical Web Made of Spider Dragline Fibers with Optical Gate Operations

• DOI: 10.1002/adom.202202563
• 发表时间: 2022-12
• 期刊: Advanced Optical Materials
• 影响因子: 9
• 作者: Hendra;Hiroshi Yamagishi;Wey Yih Heah;A. Malay;K. Numata;Yohei Yamamoto

A silk composite fiber reinforced by telechelic-type polyalanine and its strengthening mechanism

• DOI: 10.1039/d2py00030j
• 发表时间: 2022
• 期刊: Polymer Chemistry
• 影响因子: 4.6
• 作者: Jianming Chen;K. Tsuchiya;H. Masunaga;A. Malay;K. Numata