Mechanism and manipulation of growth factor receptors by bioactive peptides and AFM

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06553
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43030:Functional biochemistry-related
• Research Institution: Kanazawa University
• Principal Investigator: Sakai Katsuya 金沢大学, がん進展制御研究所, 准教授 (10523318)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 受容体 リガンド / 環状ペプチド / mRNA display / タンパク質工学 / HS-AFM / HGF-MET

Outline of Final Research Achievements

To elucidate the structural basis of growth factor receptor activation and to regulate receptor, the following studies have been conducted: 1) To analyze the structure of native active complexes of growth factor receptors that are formed on living cell membranes in very small amounts for a short time, a system to fix and purify the native complexes by chemical cross-linking was established. We are now analyzing its structure at high resolution by AFM observation and cryo-EM. 2) We have verified and developed a method to recognize unlabeled receptors in real time and in real space by using cyclic peptide coated AFM probes. 3) By presenting a receptor-binding cyclic peptide in the loop structures of the antibody Fc, we were able to create a growth factor mimetic that has a long half-life and crosses the blood-brain barrier.

Free Research Field

ケミカルバイオロジー

Academic Significance and Societal Importance of the Research Achievements

1) 本研究で確立した手法はこれまで困難であった受容体活性化複合体の構造解析を可能にし、この構造情報は創薬などに役立つと考えられる。2) タンパク質結合環状ペプチドを固定したプローブを用いたHS-AFMによって非標識タンパク質をリアルタイムかつ実空間で認識する方法は、nativeな状態で分子認識を可能にし、広範な応用が考えられる。3) 受容体結合環状ペプチドを抗体Fc内に提示することによって血中半減期が短い、脳内に到達しないといった増殖因子の欠点を克服する増殖因子ミメティックの作成を可能にした。この成果は増殖因子の治療適応を拡大する技術基盤となると考えられる。