Comparison of kinetic models for analysis of pyruvate-to-lactate exchange by hyperpolarized 13 C NMR.
Comparison of kinetic models for analysis of pyruvate-to-lactate exchange by hyperpolarized 13 C NMR.
复制标题
通过超极化 13 C NMR 分析丙酮酸与乳酸交换的动力学模型的比较。
DOI：
10.1002/nbm.2801
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发表时间：
2012-11
影响因子：
2.9
通讯作者：
Malloy, Craig R.
中科院分区：
文献类型：
--
作者：
Harrison, Crystal;Yang, Chendong;Jindal, Ashish;DeBerardinis, Ralph J.;Hooshyar, M. A.;Merritt, Matthew;Sherry, A. Dean;Malloy, Craig R.
关键词：
特异性酶催化反应的活性可以通过超极化（HP）底物的13 C NMR在体内检测。随着时间的推移，从HP基板和产品的信号，获得了一些不同的数学模型，以确定通量，但这些模型还没有进行严格的比较。在这项研究中，两个池和三个池的一阶模型被构建来测量通过乳酸脱氢酶在分离的胶质母细胞瘤细胞中的流量，通过加入超极化的[1- 13 C]丙酮酸后的乳酸和丙酮酸的NMR检测。质谱法（MS）用于独立监测13 C富集内和细胞外乳酸。六个模型进行了评估，使用时间依赖性丙酮酸C2和乳酸C1 HP NMR数据通过使用选择性激发脉冲加上13 C富集数据从细胞内和细胞外乳酸MS测量。一个三池双向模型提供了最准确的描述丙酮酸代谢在这些细胞。利用丙酮酸和乳酸的T1的计算值以及脉冲的影响，当仅HP数据可用时，通过两个池双向和单向模型很好地确定了通过乳酸脱氢酶（LDH）的初始通量。三池模型对于拟合MS和HP的组合数据是必要的，但当仅通过HP测量乳酸盐外观时，更简单的两池交换模型足以确定13 C乳酸盐浓度。
The activity of specific enzyme-catalyzed reactions may be detected in vivo by 13C NMR of hyperpolarized (HP) substrates. The signal from HP substrates and products, acquired over time, have been fit to a number of different mathematical models to determine fluxes, but these models have not been critically compared. In this study, two-pool and three-pool first-order models were constructed to measure flux through lactate dehydrogenase in isolated glioblastoma cells by NMR detection of lactate and pyruvate following addition of hyperpolarized [1-13C]pyruvate. Mass spectrometry (MS) was used to independently monitor 13C enrichment in intra- and extracellular lactate. Six models were evaluated using time dependent pyruvate C2 and lactate C1 HP NMR data acquired by use of selective excitation pulses plus 13C enrichment data from intracellular and extracellular lactate measured by MS. A three-pool bi-directional model provided the most accurate description of pyruvate metabolism in these cells. With computed values for the T1 of pyruvate and lactate as well as the effect of pulsing, the initial flux through lactate dehydrogenase (LDH) was well-determined by both the two-pool bidirectional and unidirectional models when only HP data was available. The three-pool model was necessary to fit the combined data from both MS and HP, but the simpler two-pool exchange model was sufficient to determine the 13C lactate concentration when the lactate appearance was measured only by HP.
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影响因子：
5.7
作者：
Bohndiek SE;Kettunen MI;Hu DE;Witney TH;Kennedy BW;Gallagher FA;Brindle KM
通讯作者：
Brindle KM
影响因子：
3.3
作者：
Keshari, Kayvan R.;Kurhanewicz, John;Jeffries, Rex E.;Wilson, David M.;Dewar, Brian J.;Van Criekinge, Mark;Zierhut, Matthew;Vigneron, Daniel B.;Macdonald, Jeffrey M.
通讯作者：
Macdonald, Jeffrey M.
影响因子：
3.3
作者：
Spielman, Daniel M.;Mayer, Dirk;Pfefferbaum, Adolf
通讯作者：
Pfefferbaum, Adolf
影响因子：
3.3
作者：
Chen, Albert P.;Albers, Mark J.;Vigneron, Daniel B.
通讯作者：
Vigneron, Daniel B.
影响因子：
6.3
作者：
MASON, GF;GRUETTER, R;NOVOTNY, EJ
通讯作者：
NOVOTNY, EJ