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[2,3-(13)C]琥珀酸二甲酯在大鼠肝细胞中的代谢

Metabolism of the dimethyl ester of [2,3-(13)C]succinic acid in rat hepatocytes.

作者信息

Malaisse W J, Ladrière L, Jijakli H, Laatikainen R, Niemitz M, Verbruggen I, Biesernans M, Willem R

机构信息

Laboratory of Experimental Medicine, Brussels Free University (ULB), Belgium.

出版信息

Mol Cell Biochem. 1998 Dec;189(1-2):137-44. doi: 10.1023/a:1006993629790.

Abstract

Hepatocytes prepared from overnight fasted rats were incubated for 120 min in the presence of the dimethyl ester of [2,3-(13)C]succinic acid (10 mM). The identification and quantification of 13C-enriched metabolites in the incubation medium were performed by a novel computational strategy for the deconvolution of NMR spectra with multiplet structures and constraints. The generation of 13C-labelled metabolites, including succinate, fumarate, malate, lactate, alanine, aspartate and glucose accounted for about half of the initial amount of the ester present in the incubation medium. A fair correlation was observed between the experimental abundance of each 13C-labelled glucose isotopomer and the corresponding values derived from a model for the metabolism of [2,3-(13)C]succinate. Newly formed glucose was more efficiently labelled in the carbon C5 than C2, as well as the carbon C6 than C1, supporting the concept that D-glyceraldehyde-3-phosphate may undergo enzyme-to-enzyme channelling between glyceraldehyde-3-phosphate dehydrogenase and phosphofructoaldolase.

摘要

从禁食过夜的大鼠制备的肝细胞,在[2,3-(13)C]琥珀酸二甲酯(10 mM)存在的情况下孵育120分钟。通过一种用于具有多重结构和约束的核磁共振谱去卷积的新型计算策略,对孵育培养基中富含13C的代谢物进行鉴定和定量。13C标记的代谢物的生成,包括琥珀酸、富马酸、苹果酸、乳酸、丙氨酸、天冬氨酸和葡萄糖,约占孵育培养基中酯初始量的一半。在每个13C标记的葡萄糖同位素异构体的实验丰度与源自[2,3-(13)C]琥珀酸代谢模型的相应值之间观察到良好的相关性。新形成的葡萄糖在碳C5比在碳C2、以及在碳C6比在碳C1上被更有效地标记,这支持了3-磷酸甘油醛可能在3-磷酸甘油醛脱氢酶和磷酸果糖醛缩酶之间进行酶到酶通道转运的概念。

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