生体膜機能分子のリン脂質平面膜法による解析

1991 Fiscal Year Annual Research Report

• Project/Area Number: 02304067
• Research Institution: Himeji Institute of Technology, Faculty of Science
• Principal Investigator: 平田 肇 姫路工業大学, 理学部, 教授 (40049052)
• Co-Investigator(Kenkyū-buntansha): 高木 雅行 大阪大学, 教養部, 助教授 (20028145) ; 曽我部 正博 名古屋大学, 医学部, 助教授 (10093428) ; 津田 基之 姫路工業大学, 理学部, 教授 (60045458) ; 桐野 豊 九州大学, 薬学部, 教授 (10012668) ; 葛西 道生 大阪大学, 基礎工学部, 教授 (40022595)
• Keywords: リン脂質平面膜 / イオンチャンネル / イオンポンプ / 情報伝達 / 生体膜機能素子 / 合成ペプチド / クライオ電子顕微鏡 / コンピュ-タ-シミュレ-ション

Research Abstract

本総合研究の本年度の主な成果は次の通りである。
1)シナプス小胞膜に存在するC1^~ーチャネルを平面膜法で解析したところ少なくとも2種類の性質の異なるものが同定された。さらにシナプス小胞とリポソ-ムとの融合活性に蛋白性因子が関与することが示された(曽我部、葛西)。
2)ポリエ-テル及び脂肪酸の4級アミン化合物との混合によりK^+イオンに特異性のあるチャネルが形成されることが示された(曽我部)。また、イオンチャネルのモデルとしてグラミシジンや合成ペプチドを用い、それらの化学修飾とイオン透過との関連が詳細に検討された(老木、木下)。 3)インフルエンザウィルスの膜融合モチ-フであるペプチドを化学合成し、類縁ペプチドによる膜融合の機序についての解明がなされた(高橋)。 4)イオンチャネルの精製法として平面膜法を用い、筋小胞体からK^+及びCl^~チャネルの精製に成功した(葛西)。 5)凍結・融解、脱水・水和、PEG法などにより巨大リポソ-ムを作製し、再構成したイオンチャネルの性質について平面膜法との比較が詳細に検討された(桐野)。 6)新たな平面膜法としてpinching oil layer methodが考案され、再現性のよいイオンチャネル活性測定法が開発された(高木)。7)無脊椎動物(タコ)の視細胞における光情報伝達に関与するロドプシンやGー蛋白質、cGMP依存性カチオンチャネルに関する詳細な解析がなされた(津田、平田)。 8)バクテリオロドプシンやH^+ーATPaseなどのイオンポンプを直接挿入法あるいは膜融合ペプチド法など、効率よく平面膜に再構成する方法が開発された(宗行、平田)。 9)電子顕微鏡を用いてCl^~チャネルなどの形態が詳しく計測され、イオン透過路などの微細構造に関しての推定が行われた(藤墳)。 10)時間分解能クライオ電子顕微鏡技術が考案され、5ミリ秒の時間分解能で蛋白質の構造変化の解析が行われた(徳永)。 11)コンピュ-タシミュレ-ションによるリン脂質2分子層膜の中でのリン脂質分子の動きを可視化し、その拡散速度や酸素分子の透過速度の推定が行われた(楠見)。

Research Products

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