Functional analysis of Tmem86a using oligodendorocyte progenitor cells

2021 Fiscal Year Final Research Report

• Project/Area Number: 18K06000
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 42020:Veterinary medical science-related
• Research Institution: Osaka Prefecture University
• Principal Investigator: Komori Masayuki 大阪府立大学, 生命環境科学研究科, 教授 (40183347)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Keywords: プラスマローゲン / Tmem86a / リゾプラスマロゲナーゼ / オリゴデンドロサイト前駆細胞

Outline of Final Research Achievements

Since the experiments using oligodendrocyte progenitor cells was not successful, it was decided to perform functional analysis of Tmem86a using a heterologous expression system of FLAG-tagged human Tmem86a. Although heterogeneous expression of FLAG tagged human Tmem86a alone was not successful, four chimeric proteins (Ch1 to 4) fused so that the N-terminal side is Tmem86b and the C-terminal side is Tmem86a, were possible to express them in methylotrophic yeast. In particular, Ch4 includes a nearly full-length Tmem86a. In addition, the expression of TMEM86a mRNA was stronger in rat brains than in other tissues, suggesting that Tmem86a may play an important role in the brain.

Free Research Field

生化学、分子生物学、獣医学

Academic Significance and Societal Importance of the Research Achievements

FLAGタグ付加ヒトTmem86bを異種過剰発現する酵母ミクロソームから可溶化・精製した標品を用いてリポソーム再構成系でのリゾプラスマロゲナーゼ活性を測定したが検出できず、Tmem86bがリゾプラスマロゲナーゼを持たないというごく最近の報告(文献3)を支持する結果が得られた。一方、FLAGタグ付加ヒトTmem86bとTmem86aのキメラタンパク質のうち、ほぼ全長のTmem86aを含むCh4についてメタノール資化酵母を用いる異種発現系の確立に成功したことは、Tmem86aの生化学的な機能解析などを行う上で学術的に有用である。