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Nucleoside Triphosphates - From Synthesis to Biochemical Characterization
Published on: April 3, 2014
放射性合成共因子:同型,同機能,および反応性NAD+アナログ
Alexander R Rovira1, Andrea Fin1, Yitzhak Tor1
1Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093-0358, United States.
Journal of the American Chemical Society
|October 19, 2017
まとめ
新しい光NAD+アナログであるNtzAD+が合成され,重要な酵素の活力のある基板であることが実証された. 酵素反応中に光が変化し,ネイティブのNAD+とは異なり,リアルタイムでモニタリングが可能です.
科学分野:
- 生物化学
- 化学生物学
- 生物物理化学
背景:
- ニコチナミドアデニンジヌクレオチド (NAD+) は,多くの代謝過程における重要なコファクターである.
- NAD+とNADHの非光性により,NAD+に依存した酵素反応をリアルタイムで監視することは困難です.
研究 の 目的:
- 生物化学的応用のための光NAD+アナログであるNtzAD+を合成し,特徴づけること.
- 酵素反応における基質としてのNtzAD+の有用性を評価し,その光物理的性質を評価する.
- 光スペクトロスコーピーを用いてNtzAD+を含む酵素変異のリアルタイムモニタリングを実証する.
主な方法:
- イソチアゾール[4,3-d]ピリミジンコアベースの光NAD+アナログ (NtzAD+) の合成.
- イーストアルコール脱水素酵素と乳酸脱水素酵素をNtzAD+で用いた酵素分析
- NtzAD+およびその縮小形 (NtzADH) の光物理的特徴
- NADaseとリボシルトランスファーゼ (ART5,CTA) の基板評価
主要な成果:
- NtzAD+とNtzADHはアルコールと乳酸脱水酵素の基質として機能し,ネイティブのNAD+/NADHと同等の反応率を有する.
- 光の減少は,NtzAD+からNtzADHへの変換に伴い,NAD+/NADHの振る舞いを反映しています.
- NtzAD+はNADaseとリボシルトランスファーゼ (ART5,CTA) によって処理され,tzADP-リボースを生成する.
- NtzAD+を含む酵素反応は,光スペクトロスコーピーで容易に監視できます.
結論:
- NtzAD+は,生化学研究に適したNAD+の機能的な光アナログです.
- NtzAD+/NtzADHの補完的な光物理的行動は,酵素反応のリアルタイム光モニタリングを可能にします.
- NtzAD+は,NAD+に依存する酵素と経路を研究するために,非光NAD+よりも重要な利点を提供します.
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