フォトアフィニティラベリングと質量スペクトロメトリーベースのプロテオミクスによるFKBP12-ラパミシン-FRBの結合サイトホットスポットマップ
Hope A Flaxman1, Chia-Fu Chang1, Hung-Yi Wu1
1Department of Chemistry and Chemical Biology , Harvard University , 12 Oxford Street , Cambridge , Massachusetts 02138 , United States.
Journal of the American Chemical Society
|July 17, 2019
まとめ
この研究は,ラパミシンのような複雑な分子のための結合部位をマッピングするために,光相関性ラベリング (PAL) と質量スペクトロメトリ (MS) を拡張します. この方法は,タンパク質に新しい結合モードと潜在的な標的を明らかにします.
科学分野:
- 化学生物学
- プロテオミクス
- 構造生物学
背景:
- 質量スペクトロメトリー (MS) と組み合わせたフォトアフィニティラベリング (PAL) は,小分子結合部位の構造的特徴付けを可能にします.
- 現在の方法は単純な小分子に限られており 複雑な化合物の研究を妨げています
- 複雑な分子相互作用を理解することは 薬の発見と標的の特定に不可欠です
研究 の 目的:
- 複合分子,特に免疫抑制剤ラパミシンの結合部位をマッピングするためのPALとMSの方法を拡張する.
- プロテオーム内のマクロサイクル自然産物の新しい結合モードを構造的に定義する.
- PALデータを解釈するための正確な構造測定を抽出します.
主な方法:
- PALのためのダイアジリングループを持つフォトラパミシンプローブの開発.
- FKBP12-フォトラパミシン-FRB三重複合体の形成と分析
- 結合部位を特定するために,光照射された,消化された複合体の質量スペクトロメトリーベースの分析.
主要な成果:
- 複合的なマクロサイクルの天然製品であるラパミシンの結合部位をマッピングするためにPALとMSを成功裏に適用しました.
- 光ラパミシンによるFKBP12とFRBタンパク質の特定の結合部位の特定
- 分子モデリングにより,フォトラパミシン探査機の距離制限とラベリング半径が提供されました.
結論:
- 結合地ホットスポットマッピングは,複雑なマクロサイクル自然製品にうまく適用できます.
- 開発された方法は,PALデータを解釈するための正確な構造測定を提供します.
- このアプローチにより",薬効性のない"タンパク質標的の新しい結合部位を発見することが可能になる.
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