A Method for Multiplexed Measurement of Mitochondrial Pyruvate Carrier Activity.

The Journal of Biological Chemistry
Lawrence R GrayEric B Taylor

Abstract

The discovery that theMPC1andMPC2genes encode the protein components of the mitochondrial pyruvate carrier (MPC) has invigorated studies of mitochondrial pyruvate transport and its regulation in normal and disease states. Indeed, recent reports have demonstrated MPC involvement in the control of cell fate in cancer and gluconeogenesis in models of type 2 diabetes. Biochemical measurements of MPC activity are foundational for understanding the role of pyruvate transport in health and disease. We developed a 96-well scaled method of [(14)C]pyruvate uptake that markedly decreases sample requirements and increases throughput relative to previous techniques. This method was applied to determine the mouse liver MPCKm(28.0 ± 3.9 μm) andVmax(1.08 ± 0.05 nmol/min/mg), which have not previously been reported.KmandVmaxof the rat liver MPC were found to be 71.2 ± 17 μmand 1.42 ± 0.14 nmol/min/mg, respectively. Additionally, we performed parallel pyruvate uptake and oxidation experiments with the same biological samples and show differential results in response to fasting, demonstrating the continued importance of a direct MPC activity assay. We expect this method will be of value for understanding the contribution of the MPC activity to he...Continue Reading

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Citations

Jan 25, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Bolajoko Idiat OgunyinkaAbidemi Paul Kappo
Apr 14, 2019·The EMBO Journal·Sotiria TavoulariEdmund Rs Kunji
Nov 25, 2020·Biology·José Edwin Neciosup QuesñayAndre Luis Berteli Ambrosio
Nov 27, 2020·Brain, Behavior, and Immunity·Natália C GuimarãesFabiane H Veiga-Souza

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