Solution NMR study of low-affinity protein complexes involved in interorganelle contacts

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K06047
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43020:Structural biochemistry-related
• Research Institution: Osaka University
• Principal Investigator: Furuita Kyoko 大阪大学, 蛋白質研究所, 特任助教(常勤) (30727665)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 溶液NMR / 膜コンタクトサイト / VAP / FFAT-likeモチーフ / 化学シフト摂動実験

Outline of Final Research Achievements

The ER membrane protein VAP is known to bind to a variety of amino acid sequences called FFAT-like motifs. This interaction is involved in the formation and function of membrane contact sites between the ER and other organelles. In this study, we analyzed the interactions between VAP and nine different FFAT-like motifs using solution NMR. The results showed that the FFAT-like motifs bind to the FFAT motif-binding site of VAP with dissociation constants in the millimolar range. It was found that the phenylalanine and alanine residues, which bind to the hydrophobic pocket of VAP, are important for the binding of FFAT-like motifs. Additionally, we developed software to analyze small chemical shift perturbations.

Free Research Field

構造生物科学

Academic Significance and Societal Importance of the Research Achievements

VAPとFFAT-likeモチーフとの相互作用は、膜コンタクトサイトにおいてオートファジーや神経伝達などの重要な細胞機能に寄与し、筋萎縮性側索硬化症やパーキンソン病との関連が指摘されている。またウイルスRNAの複製にも関わっている。本研究によりこのような重要な相互作用について結合様式や親和性などの結合の詳細が明らかになった。この情報はVAPとFFAT-likeモチーフの相互作用が関わる細胞機能や疾患の解析に利用しうる。また開発した化学シフト摂動実験の解析ソフトウェアはさまざまな低親和性の相互作用解析に利用できる。