ペプチドとタンパク質のチロシンタグのチロシナーゼ媒介酸化結合
Alan M Marmelstein1, Marco J Lobba1, Casey S Mogilevsky1
1Department of Chemistry, University of California at Berkeley, Berkeley, California 94720, United States.
Journal of the American Chemical Society
|February 26, 2020
まとめ
この研究は酸化結合化学を拡張して タンパク質のチロシン残基を活性化します この方法は,免疫標識などのアプリケーションのために,タンパク質のN-およびC-端末のサイト選択的修正を可能にします.
科学分野:
- 生物結合化学
- タンパク質工学
- 酵素による変化
背景:
- *o*キノンの中間物質による酸化結合 (OC) は,効率的で部位選択的なタンパク質改変を提供します.
- チロシナーゼ媒介によるフェノールタグの分子の酸化は,温和な条件下で便利な*o*キノン生成方法を提供します.
研究 の 目的:
- ペプチドとタンパク質基板のチロシン残基を活性化するためのOC化学を拡張する.
- N端とC端の両方で変更を可能にし,コップリングパートナーオプションを拡張します.
- 新種の細菌のチロシナーゼを導入し 基板活性化を強化する
主な方法:
- チロシナーゼ酵素を用いてチロシン残基を活性化して*o*キノン電ophilesを生成する.
- 二次アミンとアニリン誘導体の核愛結合パートナーとしての評価
- 抗体 scFv と タンパク質 L C- 末端を修正する戦略の適用
主要な成果:
- タンパク質のN端とC端のチロシン残基のOC改変が成功していることが実証された.
- 二次アミンとアニリンを含む有効な結合パートナーを特定した.
- 免疫標識剤として改良されたタンパク質LとscFvの有用性を示した.
結論:
- タイロシンベースのOCは,端末でタンパク質の改変のための多用途のプラットフォームを提供します.
- 開発された方法は,タンパク質工学のための生物結合戦略を拡張します.
- 改造されたタンパク質は,免疫標識の応用で有望である.
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