C-13 NMR ISOTOPOMER AND COMPUTER-SIMULATION STUDIES OF THE NONOXIDATIVE PENTOSE-PHOSPHATE PATHWAY OF HUMAN ERYTHROCYTES
C-13 NMR ISOTOPOMER AND COMPUTER-SIMULATION STUDIES OF THE NONOXIDATIVE PENTOSE-PHOSPHATE PATHWAY OF HUMAN ERYTHROCYTES
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DOI：
10.1042/bj2960379
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发表时间：
1993-12-01
影响因子：
4.1
通讯作者：
KUCHEL, PW
中科院分区：
文献类型：
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作者：
BERTHON, HA;BUBB, WA;KUCHEL, PW
C-13双量子过滤相关光谱（DQF-COSY）为碳键断裂反应的检测提供了一种新的方法。我们报告使用这种技术来研究非氧化戊糖磷酸途径在人红细胞中的同位素交换反应。这些交换反应导致葡萄糖6-磷酸的一系列同位素异构体（同位素异构体）与果糖1,6-二磷酸和溶血物的普遍c -13标记和未标记的葡萄糖6-磷酸的混合物孵育后形成。这些同位素体在脱蛋白样品的DQF-COSY光谱中的交叉峰耦合模式中被检测到。我们已经建立了一个计算机模型，充分描述了人类红细胞中非氧化戊糖磷酸途径的反应，并在实验室用P-31 n.m.r.时间过程数据进行了测试。该模型使用碳-13核磁共振时程数据进行了改进，并扩展到包括可能由非氧化戊糖磷酸途径实验反应形成的同位素的范围。从DQF-COSY光谱中得到的同位素比与该模型的模拟结果一致。
C-13 double-quantum filtered correlation spectroscopy (DQF-COSY) provides a novel method for the detection of reactions involving carbon-bond scissions. We report the use of this technique to investigate isotopic exchange reactions of the nonoxidative pentose phosphate pathway in human erythrocytes. These exchange reactions resulted in the formation of a range of isotopic isomers (isotopomers) of glucose 6-phosphate after incubation of a mixture of universally C-13-labelled and unlabelled glucose 6-phosphate with fructose 1,6-bisphosphate and haemolysates. These isotopomers were detected in the coupling patterns of cross-peaks within the DQF-COSY spectrum of the deproteinized sample. A computer model which fully describes the reactions of the non-oxidative pentose phosphate pathway in human erythrocytes has previously been constructed and tested with P-31 n.m.r. time-course data in our laboratory. This model was refined using C-13 n.m.r. time-course data and extended to include the range of isotopomers which may be formed experimentally by the reactions of the non-oxidative pentose phosphate pathway. The isotopomer ratios obtained experimentally from the DQF-COSY spectrum were consistent with simulations generated by this model.