アルキンタグSERS タンパク質における小分子結合部位のスクリーニングと識別
Jun Ando1,2,3,4, Miwako Asanuma1,2,3, Kosuke Dodo1,2,3
1AMED-CREST, Japan Agency for Medical Research and Development , Saitama 351-0198, Japan.
Journal of the American Chemical Society
|October 11, 2016
まとめ
アルキネタグラーマンスクリーニング (ATRaS) は,タンパク質の小分子結合部位を特定します. この繊細な方法は 薬の発見とプロテオミクスのために ラマン分散と質量スペクトロメトリーを使用しています
科学分野:
- 化学生物学
- プロテオミクス
- 薬物の発見
背景:
- タンパク質の小さな分子結合部位を特定することは 薬の発見とタンパク質の機能の理解に不可欠です
- 液体染色体質スペクトロメトリー (LC-MS) を使用する現在の方法は,しばしば複雑なペプチド混合物によって制限されます.
- 改造されたアミノ酸は特定できますが 効率は難しいです
研究 の 目的:
- タンパク質の小さな分子結合部位を特定するための新しい,非常に敏感な方法を開発する.
- 複雑なペプチド混合物を分析する既存の技術の限界を克服する.
- 薬剤発見とプロテオミクスのためのLC-MS分析の選択性と効率性を高める.
主な方法:
- アルキンタグによるラマンスクリーニング (ATRaS) を開発し,アルキンタグによるタンパク質との共性結合を形成する小分子を使用した.
- 表面強化ラーマン分散 (SERS) を使って,タンパク質分解とHPLC後にアルキンタグペプチドを感知する.
- 結合場所の決定的な識別のために使用されたMS/MS.
主要な成果:
- ATRaSはペプチドスクリーニングで高い感度 (∼100フェムトモルの検出限界) と再現性を示した.
- 自動化されたATRaSシステムを用いて,システインプロテアースキャセプシンBの阻害剤結合部位を成功裏に特定した.
- トリプシンで消化された細胞溶液のような複雑な混合物でATRaSシステムの有効性を検証した.
結論:
- ATRaS技術は高分子選択性を提供し,LC-MS分析を大幅に改善します.
- この方法は,薬剤発見,プロテオミクス,メタボロミクス,および化学生物学において幅広い潜在的な応用がある.
- ATRaSは,タンパク質とリガンドの相互作用を特定するための敏感で再現可能なアプローチを提供します.
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