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脂肪和NADPH的乳标记的化学基础

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

概括

在新陈代谢研究中,的追踪因水和NADPH之间的酶催化H-D交换而复杂化. 纠正这种交换对于准确跟踪氧化还原反应和使用化水的脂肪酸合成至关重要.

科学领域:

代谢工程是代谢工程.
生物化学生物化学
系统生物学系统生物学

背景情况:

像13C和这样的同位素标记物对于研究细胞代谢至关重要.
追踪提供了对氧化还原反应的洞察力,但在准确量化NADPH生产方面面临着挑战.
在代谢研究中,NADPH中原子的命运尚未完全理解.

研究的目的:

在追踪研究中调查NADPH中未计数的的来源.
阐明水和NADPH之间酶催化H-D交换的作用.
为了澄清从D2O转化为脂肪酸的子子的途径,并使精确的代谢流量分析成为可能.

主要方法:

使用Flavin酶研究了水和NADPH之间的酶催化H-D交换.
在细胞系统中分析了NADPH的产生和的结合.
在D2O的存在下合成的脂肪酸中的量化化.

主要成果:

确定了水和NADPH之间的酶催化H-D交换作为缺少的来源.
证明孤立的NADPH不会与水交换H-D,但Flavin酶可催化细胞中的快速交换.
确定D2O主要通过NADPH标记脂肪酸,从而可以无参数计算质量同位素分布.

结论:

必须考虑水和NADPH之间的酶催化H-D交换,以准确评估NADPH的生物来源.
了解这种交换机制对于精确解释对氧化还原因子和脂肪酸代谢中的追踪研究至关重要.
这种知识允许从D2O标记实验中准确,无模型计算脂肪酸质量同位素分布.

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