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構造的に異なったペプチドおよびタンパク質におけるチロシン残基の選択的修正のための酵素カスケード

Anna-Winona Struck¹, Matthew R Bennett¹, Sarah A Shepherd¹

¹School of Chemistry and Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester M1 7DN, United Kingdom.

Journal of the American Chemical Society

|February 12, 2016

まとめ

この研究は,バイオ分子におけるフェノル基の選択的改変のために,チロシナーゼとカテキル-O-メチルトランスフェラーゼ (COMT) を用いた新しいバイオオートホーゴン化学法を導入し,ラベリングと誘導のための柔軟性を向上させています.

科学分野:

生物化学
化学生物学
分子生物学

背景:

バイオオートゴーナル化学は,様々な用途のためにバイオ分子を選択的に改造することを可能にします.
メチルトランスフェラーゼ酵素 (MTase) はラベルに用いられるが,基板特異性は限られている.
複雑なバイオ分子におけるフェノール分子の選択的誘導は依然として課題である.

研究の目的:

複雑なバイオ分子におけるフェノール分子の選択的誘導のためのより柔軟な方法論を開発する.
フェノル基の連続的な水酸化とO-アルキル化のためのタンデム酵素反応を導入する.
新しい方法の汎用性と地域選択性を実証する.

主な方法:

菌類のチロシナーゼと哺乳類のカテキルOメチルトランスフェラーゼ (COMT) を含むタンデム酵素反応を用いた.
ティロシナゼはフェノール基を中間カテコールに水酸化する.
その後,COMTはカテキル分子をO-アルキル化し,選択的改変を達成します.

主要な成果:

チロシナーゼとCOMTの組み合わせは,ペプチドとタンパク質のチロシン残基の選択的アルコキシル化を可能にします.
この方法はペプチド配列の変動に対して高い耐性を示しています.
ペプチド薬,ホルモン,サイクルペプチド抗生物質を含む様々な基質を成功裏に改造した.

結論:

タイロシナーゼ-COMTシステムは,フェノール分子を改変するための多用途で地域選択的なアプローチを提供します.
この方法は,従来のMTasesの基板特異性の制限を克服します.
様々な生物学的応用における生物分子の誘導,ラベル付け,不動化のための強力なツールを提供します.

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