非ペプチド折り合いの螺旋領域を含む複合亜鉛指の設計と構造の決定
Caterina Maria Lombardo1, Vasantha Kumar M V2, Céline Douat1
1Univ. Bordeaux, CNRS, CBMN, UMR 5248, Institut Européen de Chimie et Biologie , 2 rue Robert Escarpit , 33607 Pessac , France.
Journal of the American Chemical Society
|February 5, 2019
まとめ
研究者達は 亜鉛指の天然のアルファヘリクスを 合成の折りたたみ器に置き換えて 人工タンパク質を作りました この合成されたタンパク質ハイブリッドは 亜鉛の調整とDNAの結合を保ち 新しいバイオマテリアルへの道を開きます
科学分野:
- 生物化学
- 合成生物学
- タンパク質工学
背景:
- 折りたたみ物質はタンパク質の二次構造を模倣し,特定のタンパク質の相互作用のための合成オリゴマーの設計を可能にします.
- 合成の背骨はタンパク質の天然のペプチドセグメントの代わりになり,新しい機能を持つ人工タンパク質を潜在的に作り出すことができます.
- オリゴウレア2. 5ヘリクスは,ペプチドアルファヘリクスの構造的な類似性を共有し,三次構造を維持しながら,骨幹の置換を示唆しています.
研究 の 目的:
- Cys2His2亜鉛指のアルファヘリクセグメントをオリゴウレア折りたたみ器に置き換えて複合タンパク質を作成する.
- エンジニアリングされた亜鉛指のハイブリッドが亜鉛結合能力と原生構造を維持しているかどうかを評価する.
- 改造された亜鉛指のDNA結合活性を評価する
主な方法:
- Zif268亜鉛指3のネイティブアルファヘリクスを,イミダゾールを含むオリゴウレア配列に置き換える.
- 本来の状態の分析のために,スペクトロスコーピック技術と質量スペクトロメトリーを使用する.
- 詳細な構造分析のために核磁気共振 (NMR) スペクトロスコーピーを使用します.
- Egr1の標的DNA配列で定位によるDNA結合測定を行う.
主要な成果:
- ペプチド/オリゴウレアハイブリッドは,フォルドーマー置換による影響を受けず,成功裏に亜鉛イオンを調整した.
- NMR分析では 工学的に作られた亜鉛指は 折りたたまれた構造を 採用したことが確認されました
- ネイティブのベータシート配列とDNA結合側鎖の方向性は保存された.
- 改造された亜鉛指は,野生型に似た GC 塩基への結合を示した.
結論:
- 亜鉛指のアルファヘリクセグメントをオリゴウレア折りたたみ器に置き換えることは可能である.
- 合成された複合タンパク質は,亜鉛の調整とDNA結合を含む重要な機能を保持しています.
- この研究は,設計された折りたたみコンポーネントで人工タンパク質を構築するための重要なステップを表しています.
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