マルチプル・クリック・セレクティブtRNA合成酵素が哺乳類の細胞特有のタンパク質を拡張する
Andrew C Yang, Haley du Bois, Niclas Olsson
1Center for Tissue Regeneration, Repair and Restoration , V.A. Palo Alto Healthcare System , Palo Alto , California 94304 , United States.
Journal of the American Chemical Society
|May 19, 2018
まとめ
研究者は哺乳類の細胞に正確なラベルを付けるため,新しいバイオオートホーゴナルツールを開発し,チロシルとフェニラニルtRNA合成体を設計した. これらのツールは,より深い生物学的な理解のために,高度な,細胞型特異なプロテオミクスを可能にします.
科学分野:
- 生物化学
- 分子生物学
- プロテオミクス
背景:
- 細胞型特異のプロテオミクスは,多細胞生物の複雑な生物学的プロセスを理解するために不可欠です.
- 哺乳類のシステムでは既存のバイオオートゴーナルツールには限界があります.
研究 の 目的:
- 哺乳類のバイオオルトゴナルラベリングのための新しいアミノアシル-tRNA合成酵素を開発する.
- 生きた哺乳類の細胞型特異なプロテオミクスを可能にします
主な方法:
- タイロシル (ScTyrY43G) とフェニララニル (MmPheT413G) tRNA合成酵素を設計した.
- アジドを含む非正規のアミノ酸を新生タンパク質に組み込む.
- アジド-アルキンサイクル添加物は,フッ素素または親和樹脂で化学選択的にタグ付けするために使用されます.
主要な成果:
- ScTyrY43GとMmPheT413Gは,ヒト,ハムスター,マウスの細胞系でタンパク質を成功裏に標識しました.
- 変異合成酵素は,正規の10倍以上のアジドを含むアミノ酸を選択的に活性化します.
- マウスにおけるメラノーマ腫瘍プロテオームとプラズマシクレトームのインビボラベリングが達成された.
結論:
- エンジニアリングされた合成酵素は,哺乳類の残基特有のバイオオルトゴン化学のための新しいツールを提供します.
- これらのツールは,生きた哺乳類におけるより堅牢で包括的な細胞型特異のプロテオミクスを促進します.
- プロテオームの高度なイメージングと質量スペクトロメトリックの特徴づけを可能にします.
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