Development and AFM observation of immobilized planar membranes with controlled orientation of membrane proteins and asymmetry of lipid membranes

2021 Fiscal Year Final Research Report

• Project/Area Number: 18K19422
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 48:Biomedical structure and function and related fields
• Research Institution: Kanazawa University
• Principal Investigator: Sumino Ayumi 金沢大学, ナノ生命科学研究所, 助教 (80717140)
• Project Period (FY): 2018-06-29 – 2022-03-31
• Keywords: 高速原子間力顕微鏡 / 非対称膜

Outline of Final Research Achievements

The purpose of this study is to develop a method to prepare supported lipid bilayers with simultaneous control of membrane protein orientation and lipid membrane asymmetry, and to observe the dynamics of membrane proteins whose function is related to lipid membrane asymmetry by high-speed AFM. By FY 2019, planar membranes were prepared on AFM substrates by the droplet contact method. Using a micromanipulator and a microinjector on the sample stage of a tip-scanning high-speed AFM, we were able to prepare a uniform lipid film on the AFM substrate. However, there was a problem that the optical system of the tip-scanning high-speed AFM was unstable. Therefore, in FY2021, we replaced the tip-scanning high-speed AFM with a more stable sample-scanning high-speed AFM and adjusted it to a level where it could operate stably.

Free Research Field

生物物理学

Academic Significance and Societal Importance of the Research Achievements

AFM基板上での膜タンパク質の配向と脂質膜の非対称性の同時制御ができれば、膜タンパク質の機能動態に及ぼす脂質の非対称性の影響などについて分子レベルでの議論が可能になる。