Comprehensive Single-Molecule Protein Quantification Analysis Using Ribosome-Integrated Nanopore Technology

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K18232
• Research Category: Grant-in-Aid for Challenging Research (Pioneering)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
• Research Institution: The University of Tokyo
• Principal Investigator: UEMURA Sotaro 東京大学, 大学院理学系研究科(理学部), 教授 (00447442)
• Project Period (FY): 2021-07-09 – 2024-03-31
• Keywords: １分子計測 / ナノポア計測 / ペプチド / 機械学習

Outline of Final Research Achievements

In this study, the goal was to develop a foundational technology for comprehensive protein analysis using ribosome-integrated nanopores to advance the comprehensive analysis of peptide molecules. Throughout the duration of the study, we first selected the translocon SecYEG and the molecular motor SecA complex from among the membrane channel proteins available for nanopore measurements, and successfully reconstituted them on artificial lipid bilayers. As a result, we detected membrane insertion current signals and were able to observe characteristic nanopore current waveforms only in the presence of peptide molecule substrates and ATP. Currently, we are attempting to detect current signal changes in response to artificial peptides containing repetitive amino acid sequences.

Free Research Field

生物物理学

Academic Significance and Societal Importance of the Research Achievements

本研究による１分子ペプチドのアミノ酸配列同定の学術的意義や社会的意義は極めて大きく、その効果は計り知れない。特にペプチドはそもそも分解酵素によって分解されやすく分子数が少なく、増幅する技術も存在しない。基礎研究においてはシグナル伝達、アポトーシス制御だけでなく成長因子や免疫応答、神経伝達などの細胞機能制御に関わる学問領域に大きな貢献が期待される。さらに応用研究として、抗菌ペプチド、ドラッグデリバリー、ペプチド医薬品やホルモン検出などの臨床応用や診断、創薬領域にも大きな貢献が期待される。