New methods for protein structural analysis using light-excitable transiently paramagnetic species

2019 Fiscal Year Final Research Report

• Project/Area Number: 17H03089
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Osaka University
• Principal Investigator: Matsuki Yoh 大阪大学, 蛋白質研究所, 准教授 (70551498)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 過渡的常磁性タグ / 固体NMR

Outline of Final Research Achievements

Magic-angle spinning solid-state NMR methods has recently witnessed a maturation of methodologies utilizing paramagnetic dopants for accelerating signal averaging and long-range internuclear distance measurements. These original methods had however some own limitations coming from the paramagnetism that remains during the signal acquisition period, resulting in line broadening or signal loss. In this study, light-excitable transiently paramagnetic species were explored to make a functional dopants and protein-tag whose magnetism is switchable by making on/off the excitation light. This is shown here to resolve the conventional methodological limitations, which will become a basis for new and more efficient tools for structural study of proteins.

Free Research Field

固体核磁気共鳴分光

Academic Significance and Societal Importance of the Research Achievements

本課題では励起光のオン／オフによってRFパルス列実行中とFID取り込み中で、試料に混入した分子の磁性を高速に切り換え利用する実験法を開発し、複数の機能性化合物(色素)を見出した。これは従来法の主要な欠点を克服する最初の例であり、分解能低下の心配なく感度を増強でき、また修飾残基の根元を含めた短距離から長距離情報までの核間距離情報を残さず取得できる手法の基盤となる。試料調製の面での要求も軽減するので、発現困難な試料、複合体試料の構造研究の新発展を推進する潜在能力がある、重要な基盤技術である。