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Nature Chemical Biology
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Lactate metabolism is associated with mammalian mitochondria

Nature Chemical Biologyvolume 12, pages937–943 (2016)Cite this article

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Abstract

It is well established that lactate secreted by fermenting cells can be oxidized or used as a gluconeogenic substrate by other cells and tissues. It is generally assumed, however, that within the fermenting cell itself, lactate is produced to replenish NAD+ and then is secreted. Here we explore the possibility that cytosolic lactate is metabolized by the mitochondria of fermenting mammalian cells. We found that fermenting HeLa and H460 cells utilize exogenous lactate carbon to synthesize a large percentage of their lipids. Using high-resolution mass spectrometry, we found that both13C and 2-2H labels from enriched lactate enter the mitochondria. The lactate dehydrogenase (LDH) inhibitor oxamate decreased respiration of isolated mitochondria incubated in lactate, but not of isolated mitochondria incubated in pyruvate. Additionally, transmission electron microscopy (TEM) showed that LDHB localizes to the mitochondria. Taken together, our results demonstrate a link between lactate metabolism and the mitochondria of fermenting mammalian cells.

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Figure 1: 125-MHz cross-polarization magic-angle spinning13C NMR spectra.
Figure 2: HeLa and H460 cells cultured in [13C3]lactate have labeled tricarboxylic acid (TCA) cycle intermediates.
Figure 3: Deuterium labels from [2H]lactate enter the mitochondria.
Figure 4: Lactate dehydrogenase B (LDHB) is present in HeLa mitochondria.
Figure 5: HeLa mitochondria use lactate as a carbon source to respire.

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Acknowledgements

This work was supported by funding from the National Institutes of Health grants R01 ES022181 (G.J.P.), R21 CA191097-01A1 (G.J.P.), R01 HL118639-03 (R.W.G.), R01 DK091538 (P.A.C.), and R01 EB002058 (J.S.), as well as grants from the Alfred P. Sloan Foundation (G.J.P.), the Camille & Henry Dreyfus Foundation (G.J.P.), and the Pew Scholars Program in the Biomedical Sciences (G.J.P.). We thank W. Beatty at Washington University's Molecular Microbiology Imaging Facility for acquiring the TEM images.

Author information

Authors and Affiliations

  1. Department of Chemistry, Washington University, St. Louis, Missouri, USA

    Ying-Jr Chen, Nathaniel G Mahieu, Manmilan Singh, Jacob Schaefer & Gary J Patti

  2. Department of Genetics, Washington University School of Medicine, St. Louis, Missouri, USA

    Xiaojing Huang & Stephen L Johnson

  3. Sanford Burnham Prebys Medical Discovery Institute, Orlando, Florida, USA

    Xiaojing Huang & Peter A Crawford

  4. Department of Medicine, Washington University School of Medicine, St. Louis, Missouri, USA

    Richard W Gross & Gary J Patti

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  1. Ying-Jr Chen

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Contributions

Y.-J.C. prepared samples, carried out biochemical assays, and performed LC/MS analyses. Y.-J.C., M.S., and J.S. performed NMR analyses. Y.-J.C., N.G.M., X.H., M.S., P.A.C., S.L.J., R.W.G., J.S., and G.J.P. contributed to experimental design and data interpretation. Y.-J.C., N.G.M., and G.J.P. wrote the manuscript.

Corresponding author

Correspondence toGary J Patti.

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Competing interests

G.J.P. is a scientific advisory board member for Cambridge Isotope Laboratories, Tewksbury, Massachusetts, USA. R.W.G. has financial relationships with LipoSpectrum and Platomics. P.A.C. serves as a consultant and on a scientific advisory board for Janssen Pharmaceuticals, Titusville, New Jersey, USA.

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Supplementary Results, Supplementary Figures 1–12 and Supplementary Tables 1–3. (PDF 13674 kb)

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Chen, YJ., Mahieu, N., Huang, X.et al. Lactate metabolism is associated with mammalian mitochondria.Nat Chem Biol12, 937–943 (2016). https://doi.org/10.1038/nchembio.2172

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