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脂肪とNADPHのデュテリウムラベリングの化学的基礎

Zhaoyue Zhang^1,2, Li Chen^1,2, Ling Liu^1,2

¹Lewis-Sigler Institute for Integrative Genomics, Princeton University , Princeton, New Jersey 08544, United States.

Journal of the American Chemical Society

|September 16, 2017

まとめ

代謝研究におけるデウテリウムの追跡は,水とNADPHの間の酵素触媒によるH-D交換によって複雑になる. この交換を修正することは,脱酸化水を用いて酸化還元反応と脂肪酸合成を正確に追跡するために不可欠です.

科学分野:

メタボリック・エンジニアリング
生物化学
システム生物学

背景:

13Cやデュテリウムのような同位体トレーサーは細胞代謝の研究に不可欠です
デウテリウムの追跡は,酸化還元反応の洞察を提供しているが,NADPHの生産を正確に定量化するには課題に直面している.
代謝研究中のNADPH中の水素原子の運命は,完全に理解されていません.

研究の目的:

デュテリウムトレース研究中にNADPHに含まれていない水素の源を調べる.
水とNADPH間の酵素触媒によるH-D交換の役割を明らかにする.
D2Oから脂肪酸へのデウテロンの経路を明らかにし,正確な代謝流量分析を可能にします.

主な方法:

フラビン酵素を用いた水とNADPHの酵素触媒によるH-D交換を調査した.
細胞システムにおけるNADPHの生成とデュテリウムの組み込みを分析した.
D2Oの存在で合成された脂肪酸の量化デュテレーション.

主要な成果:

水とNADPHの間の酵素触媒によるH-D交換を欠けている水素の源として特定した.
隔離されたNADPHは水とH-Dを交換しないが,フラビン酵素は細胞の急速な交換を触媒とする.
D2Oは主にNADPHを介して脂肪酸を標識し,質量同位体分布のパラメータフリー計算を可能にします.

結論:

NADPHの生物学的源を正確に評価するには,水とNADPHの間の酵素触媒によるH-D交換を考慮する必要があります.
この交換メカニズムの理解は,酸化還元因子と脂肪酸代謝におけるデウテリウム追跡研究の正確な解釈に不可欠である.
この知識は,D2Oのラベリング実験から脂肪酸の質量同位体分布の正確なモデルフリー計算を可能にします.

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