NMR approach for the functional structural analyses of human membrane proteins

2017 Fiscal Year Final Research Report

• Project/Area Number: 15H04340
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Yokohama City University
• Principal Investigator: Takahashi Hideo 横浜市立大学, 生命医科学研究科, 教授 (60265717)
• Co-Investigator(Kenkyū-buntansha): 竹内 恒 国立研究開発法人産業技術総合研究所, 生命工学領域, 研究チーム長 (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): 2015-04-01 – 2018-03-31
• Keywords: NMR / 膜タンパク質 / 酵母

Outline of Final Research Achievements

In this study, we have developed a novel and cost-effective strategy for P. pastoris expression systems to selectively incorporate 13C into methyl groups of Val and Leu by using the alpha-ketoacid precursor. Methyl-specific isotopically-labeled samples are quite pivotal for investigating the structures and interactions of large macromolecules such as membrane proteins by solution NMR. We further established an experimental procedure to prepare NLP-reconstituted human membrane protein (C99) which allows analyzing membrane proteins in lipid bilayer membranes by solution NMR. Our NMR analysis revealed that the lipid composition in the bilayer remarkably influences structure and dynamics of the transmembrane helix region.

Free Research Field

構造生物学