A 13C NMR study on fluxes into the krebs cycle of rabbit renal proximal tubular cells
A 13C NMR study on fluxes into the krebs cycle of rabbit renal proximal tubular cells
复制标题
兔肾近端小管细胞克雷布斯循环通量的 13C NMR 研究
作者：
A. Jans;D. Leibfritz
用[2‐13C]和[3‐13C]丙酮酸孵育兔肾近端小管（PCT）细胞悬液。用13C核磁共振对细胞微球的高氯酸提取物进行了检测。所有实验都表明，富集的乳酸、丙氨酸、谷氨酸和谷氨酰胺是主要的代谢中间体，而谷胱甘肽（GSH）的谷氨酸残基中有少量富集。从这些实验可以推断，PCT细胞表现出高度的糖酵解活性，而葡萄糖的富集表现为糖异生。讨论了用13C核磁共振估计丙酮酸羧化酶进入Krebs循环的通量与丙酮酸脱氢酶进入Krebs循环的通量之比。从10 mM 13C标记的丙酮酸孵育中，我们计算出谷氨酸碳原子的相对富集，在兔肾近端小管中丙酮酸羧化酶与丙酮酸脱氢酶的比值为1.44±0.04。
Suspensions of rabbit renal proximal tubular (PCT) cells were incubated with [2‐13C] and [3‐13C]pyruvate. The perchloric acid extracts of the cell pellets were examined by 13C NMR. All experiments showed that enriched lactate, alanine, glutamate, and glutamine were the main metabolic intermediates, and that enrichment to a minor extent was found in the glutamate residue of glutathione (GSH). From these experiments, it could be deduced that PCT cells show a highly glycolytic activity, whereas enrichment of glucose exhibits gluconeogenesis. The estimation by 13C NMR of the ratio of the flux into the Krebs cycle via pyruvate carboxylase to the flux via pyruvate dehydrogenase is discussed. From incubations with 10 mM 13C‐labelled pyruvate, we calculated from the relative enrichments of the glutamate carbon atoms that the ratio of pyruvate carboxylase to pyruvate dehydrogenase is 1.44 ± 0.04 in rabbit renal proximal tubules.
柠檬酸循环中同位素标记的数学分析及其在灌注大鼠心脏 13C NMR 研究中的应用。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Chance,EM;Seeholzer,SH;Kobayashi,K;Williamson,JR
通讯作者：
Williamson,JR