期刊
CELL METABOLISM
卷 22, 期 5, 页码 936-947出版社
CELL PRESS
DOI: 10.1016/j.cmet.2015.08.021
关键词
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资金
- Public Health Service [R01 AA-021984, R01 DK-092606, K08 DK-080142]
- Yale CTSA [UL1 RR-0024139]
- DRC [P30 DK-45735]
- O'Brien Kidney Center [P30 DK79310-01A1]
- Mouse Metabolic Phenotyping Center [U24 DK-059635]
- Liver Center [P30-DK-034989]
- Cancer Center [IRG-058-012-53]
Mass isotopomer multi-ordinate spectral analysis (MIMOSA) is a step-wise flux analysis platform to measure discrete glycolytic and mitochondrial metabolic rates. Importantly, direct citrate synthesis rates were obtained by deconvolving the mass spectra generated from [U-C-13(6)]-D-glucose labeling for position-specific enrichments of mitochondrial acetylCoA, oxaloacetate, and citrate. Comprehensive steady-state and dynamic analyses of key metabolic rates (pyruvate dehydrogenase, beta-oxidation, pyruvate carboxylase, isocitrate dehydrogenase, and PEP/pyruvate cycling) were calculated from the position-specific transfer of C-13 from sequential precursors to their products. Important limitations of previous techniques were identified. In INS-1 cells, citrate synthase rates correlated with both insulin secretion and oxygen consumption. Pyruvate carboxylase rates were substantially lower than previously reported but showed the highest fold change in response to glucose stimulation. In conclusion, MIMOSA measures key metabolic rates from the precursor/product position-specific transfer of C-13-label between metabolites and has broad applicability to any glucose-oxidizing cell.
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