Dynamic analysis of precursor proteins passing through the translocase of the mitochondrial outer membrane complex

2022 Fiscal Year Final Research Report

• Project/Area Number: 20H02583
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 28040:Nanobioscience-related
• Research Institution: Kanazawa University
• Principal Investigator: Araiso Yuhei 金沢大学, 保健学系, 助教 (20753726)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: ミトコンドリア / タンパク質輸送 / TOM複合体 / 膜タンパク質複合体 / 高速原子間力顕微鏡

Outline of Final Research Achievements

The TOM complex was purified from a yeast overexpressing strain. HS-AFM analysis demonstrated that the purified TOM complex forms a trimeric structure in solution and tends to dissociate into a stable core-dimer and a peripheral monomer. Furthermore, we succeeded in real-time imaging of interaction between the TOM complex and mitochondrial precursor proteins, in which a substrate protein is added to the TOM complex under HS-AFM observation. We are currently attempting to identify the substrate-binding region of the TOM complex by applying various substrates.

Free Research Field

構造生物化学、生化学、ナノバイオサイエンス

Academic Significance and Societal Importance of the Research Achievements

TOM複合体は３量体と２量体の二つの分子種が混在した不均一状態であり、TOM複合体の機能の実態に迫るには、狙ったフォームだけを取り出して１分子解析する新規手法の確立が必要である。我々はHS-AFMを用いて３量体TOM複合体の解離と基質認識機構を観察し、従来手法では解析することの難しかったTOM複合体のダイナミクスを可視化することができた。本成果を起点にTOM複合体の新たな分子機構解明が期待される。