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フラビンとニコチナミドのリドックス共因子を結合するRNAアプタマー

C T Lauhon¹, J W Szostak

¹Department of Molecular Biology, Massachusetts General Hospital, Boston 02114, USA.

Journal of the American Chemical Society

|February 1, 1995

まとめ

研究者らは,リボフラビンとニコチナミドモノヌクレオチドを結合するRNAアプタマーを開発した. これらのRNA分子は,レドックス反応に関与する新しいリボ酵素を作り出す可能性を示しており,代謝経路を模倣しています.

科学分野:

バイオケミストリーと分子生物学
合成生物学合成生物学とは
RNA アプタマーテクノロジー

背景:

リボフラビン (Rb) とベータ・ニコチナミド・モノヌクレオチド (NMN) は,多くの代謝的酸化還元反応における重要な共因子である.
これらの反応を模倣または制御するための人工システムの開発は,生化学研究の重要な分野です.
RNAアプタマーは,分子認識と触媒の汎用性のあるプラットフォームを提供します.

研究の目的:

リボフラビンとNMNに特異的に結合するRNAアプタマーを分離し,特徴づけること.
これらのアプタマーにおけるコファクター結合とリドックス特異性の構造的基礎を調査する.
これらのアプタマーモチーフが新たなリボ酵素を開発する可能性を調査する.

主な方法:

In vitro選択 (SELEX) で,RbとNMNに結合するRNAアプタマーを分離する.
DMSの保護測定は,RNAの二次構造とコファクターの相互作用を決定する.
ミュタゲネシスと再選択により,重要な結合領域を特定し,特異性を高める.
結合強度を定量化するための親和度測定 (Kd).

主要な成果:

酸化リボフラビンを結合するG-クォーテットモチーフを持つRNAアプタマーを特定し,マイクロモラー親和性を示した.
選択されたRNAアプタマーで,NADとNADHを1度以上の差で区別する.
NMN結合アプタマーの二次構造を特徴付け,複雑な幹回路相互作用を明らかにした.
NMNの結合特異性を高める特定の変異を発見した.

結論:

RNAは,RbやNMNのような重要なコファクターを高親和度で結合することができる.
RNAアプタマーは,特にニコチナミド共因子に対して,酸化還元状態の差別化のために設計することができます.
これらの発見は,新種のリボ酵素をリドックス触媒化のために設計するための基礎を提供します.

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