Investigating spider silk self-assembly via liquid-liquid phase separation
| Project/Area Number |
21K06043
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 43020:Structural biochemistry-related
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| Research Institution | Tokyo Institute of Technology (2022)
Institute of Physical and Chemical Research (2021) |
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Principal Investigator |
Malay Ali 東京工業大学, 物質理工学院, 研究員 (40467006)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Project Status |
Granted (Fiscal Year 2022)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2023: ¥260,000 (Direct Cost: ¥200,000、Indirect Cost: ¥60,000)
Fiscal Year 2022: ¥390,000 (Direct Cost: ¥300,000、Indirect Cost: ¥90,000)
Fiscal Year 2021: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
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| Keywords | silk / biomimetic / liquid phase separation / coacervate / nanofibril / self-assembly / biopolymer / biomolecular condensate / biomimetics / phase separation / spider / biomaterial / biochemistry / silk protein |
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| Outline of Research at the Start |
Spider silk is a protein biopolymer with extreme mechanical properties; however, the mechanism for silk fiber formation is not well understood. Recently, we found that spider silk undergoes self-assembly via liquid-liquid phase separation and nanoscale fibril formation in response to different chemical stimuli. In this project we will explore the sequence features of silk proteins that govern the the self-assembly process. These results will help develop methods to produce artificial spider silk fibers that mimic the hierarchical structure and mechanical properties of natural spider silk.
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| Outline of Annual Research Achievements |
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.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Much progress was achieved this year, and work is still undergoing. For instance, for the study on MaSp1 (in isolation) experimental work has been completed and the manuscript has been written, but so far it has not been successfully published yet. For the other aspects, such as the study on the synergistic effects of MaSp1/2/3 spidroins, the work has not been fully completed; much of these relate to the difficulty of optimizing the experimental conditions, but it is also related to delays related to scheduling of beam-time at SPring-8, as well as to the effect of moving to a new institution (Tokyo Institute of Technology).
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| Strategy for Future Research Activity |
In the following year, I wish to pursue the experiments related to determining the effects of the different repetitive sequence motifs of MaSp proteins on the self-assembly of recombinant spider silk, as well as on the composite nature of the structure and function of spider dragline silk, until the successful publication of results in high-impact publications.
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Report
(2 results)
Research Products
(10 results)
