非常に高い熱的および化学的な安定性を持つイコサヘドラルタンパク質ケージのコイル-コイル-メディエイト組立
Ajitha S Cristie-David1, Junjie Chen1, Derek B Nowak2
1Department of Chemistry , University of Michigan , Ann Arbor , Michigan 48109 , United States.
Journal of the American Chemical Society
|May 24, 2019
まとめ
科学者たちは,イコサエドール対称性からインスパイアされた新しいデザインを使用して,非常に安定した,空洞のタンパク質ケージを設計しました. この新しいナノスケールアーキテクチャは 生体工学と医学における 酵素の安定性を高めます
科学分野:
- 生物化学
- 合成生物学
- 構造生物学
背景:
- タンパク質の自己組み立ては 自然界において極めて重要であり 合成生物学における重要な目標です
- 酵素の安定化は 生物工学と医学において極めて重要です
研究 の 目的:
- シンメトリー指向の原理を用いて 非常に安定した 酵素活性のある ホールタンパク質ケージを設計・構築する.
- タンパク質の安定性に対する高階構造の影響を調査する.
主な方法:
- トリメアタンパク質 (TriEst) とペンタメリク・コイルドメインをグリシン・リンカーで遺伝的に融合させ,対称性指向設計を行う.
- 最適なケージ形成を特定するためのスクリーニングリンク長 (8つのグリシン残留物を持つIco8).
- ダイナミック光散射,ネガティブステイン,冷凍電子顕微鏡,原子力顕微鏡,およびIR光誘導力顕微鏡を用いた特徴付け.
主要な成果:
- 60個のサブユニット (20個のトライエストトリマー) を含む,アイコアヘドラル形状の空洞のタンパク質ケージ (Ico8) を成功裏に組み立てました.
- Ico8のケージは120°Cと8Mのグアニジン塩化物で折りたたまれた状態で,著しい安定性を示した.
- コンポーネントタンパク質 (TriEst) の安定性は著しく低かった (Tm ≈ 75 °C,グアニジン塩化物1.5Mでの変性).
結論:
- プロテインケージの高階構造は,個々のサブユニットには存在しない新興の安定性をもたらします.
- この設計されたタンパク質ケージは 頑丈で機能的なナノマテリアルを作る 強力な戦略を示しています
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