Biosensors and control of cell functions using biomolecular-recognition nanopores

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K19039
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 37:Biomolecular chemistry and related fields
• Research Institution: Gunma University
• Principal Investigator: Kamiya Koki 群馬大学, 大学院理工学府, 助教 (70612315)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Keywords: ナノポアタンパク質 / リン脂質膜 / バイオセンシング / タンパク質改変 / 無細胞タンパク質合成系

Outline of Final Research Achievements

We created a modified OmpG that expands and truncates β-hairpins, allowing the generation of small or large nanopores compared to that of wild-type (WT) OmpG nanopores. We demonstrated the detection of various structures of DNA (branched DNA) depending on the nanopore size using OmpG WT or mutated OmpG nanopores. Insights into the changes in pore diameters will be crucial to form precise pore diameters for the detection of various types of single biomolecules and for sequencing DNA, peptides, and proteins. Next, OmpG and OmpA synthesized by a cell-free synthesis system were incorporated into nano-sized liposomes. The nanopore formation in the nano-sized liposomes was confirmed using a patch-clamp method.

Free Research Field

生体関連化学

Academic Significance and Societal Importance of the Research Achievements

βストランド数を増減させることでポア内のイオンの通過には、ポア内の水と接する部分のアミノ酸残基の電荷やかさ高さが重要な役割を果たしていることが分かった。改変体OmpGなどによって、分岐の長さが異なるT字DNAの検出に成功した。しがたって、自由にポアサイズを設計できるようになると、様々な大きさの生体分子の検出が可能になる。