Fundamental research for NMR structural analysis of human membrane proteins

2014 Fiscal Year Final Research Report

• Project/Area Number: 24370048
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Yokohama City University
• Principal Investigator: TAKAHASHI Hideo 横浜市立大学, 大学院生命医科学研究科, 教授 (60265717)
• Co-Investigator(Kenkyū-buntansha): TAKEUCHI Koh 産業技術総合研究所, 創薬分子プロファイリング研究センター, 主任研究員 (20581284)
• Co-Investigator(Renkei-kenkyūsha): SAKAKURA Masayoshi 横浜市立大学, 大学院生命医科学研究科, 助教 (20334336)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: NMR / 膜タンパク質

Outline of Final Research Achievements

Human membrane proteins (hMPs) are important targets in the structural biology field because they contain a lot of drug targets. However, expression of recombinant hMPs by conventional prokaryotic cells is usually difficult. In this study, we constructed a robust strategy to express hMPs using two yeast species, P. pastoris and K. lactis. We further developed a variety of methods to isotopically label recombinant hMPs for structural studies using nuclear magnetic resonance (NMR) spectroscopy. We expressed six hMPs in the yeast membranes, and some of them were labeled with 2H, 15N, and 13C in a methyl-group selective manner. Using these samples, we could obtain molecular properties and structural information of several hMPs. The protocol established in this study is a widely applicable method and will contribute to following NMR studies of hMPs.

Free Research Field

構造生物学