Establishment of protein folding evolution using cell-free protein synthesis and peptide tags

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06519
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43020:Structural biochemistry-related
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Niwa Tatsuya 東京工業大学, 科学技術創成研究院, 助教 (50588530)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: タンパク質フォールディング / 試験管内分子進化 / 質量分析装置 / 無細胞タンパク質合成系

Outline of Final Research Achievements

To deepen our understanding of protein folding, which is still difficult to predict, we have been trying to establish an experimental system for in vitro protein folding evolution using peptide tags and a nano-LC-tandem mass spectrometer. We have designed about 100 types of quantifiable peptide tags, investigated and optimized the measurement conditions using a cell-free protein synthesis system, and attempted folding evolution experiments of model proteins but have not yet succeeded in doing so. The problems are the high cost of measurement and the inefficiency of sample preparation and measurement. Some of these problems are expected to be improved by devising experimental conditions, and we would like to challenge these problems if possible.

Free Research Field

生化学・分子生物学

Academic Significance and Societal Importance of the Research Achievements

本研究の成果によって、質量分析装置を用いたターゲットMS解析をタンパク質の分子進化系に利用できる可能性が示されたと考えている。一方で、その実現のためには測定条件や適切なペプチドタグの設計など、考慮しなければいけない点が多数あることも判明した。これらの結果は今後ターゲットMSを似たような実験系に用いる際に非常に有用な知見となるはずである。