Colocalization of MCT1, CD147, and LDH in mitochondrial inner membrane of L6 muscle cells: evidence of a mitochondrial lactate oxidation complex.

Results of previous studies suggested a role of mitochondria in intracellular and cell-cell lactate shuttles. Therefore, by using a rat-derived L6 skeletal muscle cell line and confocal laser-scanning microscopy (CLSM), we examined the cellular locations of mitochondria, lactate dehydrogenase (LDH), the lactate-pyruvate transporter MCT1, and CD147, a purported chaperone protein for MCT1. CLSM showed that LDH, MCT1, and CD147 are colocalized with the mitochondrial reticulum. Western blots showed that cytochrome oxidase (COX), NADH dehydrogenase, LDH, MCT1, and CD147 are abundant in mitochondrial fractions of L6 cells. Interactions among COX, MCT1, and CD147 in mitochondria were confirmed by immunoblotting after immunoprecipitation. These findings support the presence of a mitochondrial lactate oxidation complex associated with the COX end of the electron transport chain that might explain the oxidative catabolism of lactate and, hence, mechanism of the intracellular lactate shuttle.

[1]  G. Brooks,et al.  Immunohistochemical analysis of MCT1, MCT2 and MCT4 expression in rat plantaris muscle , 2005, The Journal of physiology.

[2]  J. Zoll,et al.  Mitochondrial tissue specificity of substrates utilization in rat cardiac and skeletal muscles , 2005, Journal of cellular physiology.

[3]  H. Daniel,et al.  Activation of mitochondrial lactate uptake by flavone induces apoptosis in human colon cancer cells , 2005, Journal of cellular physiology.

[4]  G. Brooks,et al.  MCT1 confirmed in rat striated muscle mitochondria. , 2004, Journal of applied physiology.

[5]  G. Brooks,et al.  Pyruvate shuttling during rest and exercise before and after endurance training in men. , 2004, Journal of applied physiology.

[6]  L. Gladden Lactate metabolism: a new paradigm for the third millennium , 2004, The Journal of physiology.

[7]  D. Meredith,et al.  The SLC16 gene family—from monocarboxylate transporters (MCTs) to aromatic amino acid transporters and beyond , 2004, Pflügers Archiv.

[8]  N. Philp,et al.  MCT1 and its accessory protein CD147 are differentially regulated by TSH in rat thyroid cells. , 2003, American journal of physiology. Endocrinology and metabolism.

[9]  G. Brooks,et al.  Peroxisomal membrane monocarboxylate transporters: evidence for a redox shuttle system? , 2003, Biochemical and biophysical research communications.

[10]  Hans N. Rasmussen,et al.  Lactate dehydrogenase is not a mitochondrial enzyme in human and mouse vastus lateralis muscle , 2002, The Journal of physiology.

[11]  K. Sahlin,et al.  No evidence of an intracellular lactate shuttle in rat skeletal muscle , 2002, The Journal of physiology.

[12]  G. Brooks,et al.  Changes in MCT 1, MCT 4, and LDH expression are tissue specific in rats after long-term hypobaric hypoxia. , 2002, Journal of applied physiology.

[13]  D. Meredith,et al.  Fluorescence Resonance Energy Transfer Studies on the Interaction between the Lactate Transporter MCT1 and CD147 Provide Information on the Topology and Stoichiometry of the Complex in Situ * , 2002, The Journal of Biological Chemistry.

[14]  C. Des Rosiers,et al.  Evidence of separate pathways for lactate uptake and release by the perfused rat heart. , 2001, American journal of physiology. Endocrinology and metabolism.

[15]  G. Brooks,et al.  Lactate shuttles in nature. , 2001, Biochemical Society transactions.

[16]  G. Rutter,et al.  Expression and distribution of lactate/monocarboxylate transporter isoforms in pancreatic islets and the exocrine pancreas. , 2001, Diabetes.

[17]  A. Barclay,et al.  CD147 is tightly associated with lactate transporters MCT1 and MCT4 and facilitates their cell surface expression , 2000, The EMBO journal.

[18]  G. Brooks,et al.  Endurance training, expression, and physiology of LDH, MCT1, and MCT4 in human skeletal muscle. , 2000, American journal of physiology. Endocrinology and metabolism.

[19]  G. Brooks,et al.  Cardiac and skeletal muscle mitochondria have a monocarboxylate transporter MCT1. , 1999, Journal of applied physiology.

[20]  G. Brooks,et al.  Active muscle and whole body lactate kinetics after endurance training in men. , 1999, Journal of applied physiology.

[21]  N. Price,et al.  The proton-linked monocarboxylate transporter (MCT) family: structure, function and regulation. , 1999, The Biochemical journal.

[22]  A. Bröer,et al.  Characterization of the high-affinity monocarboxylate transporter MCT2 in Xenopus laevis oocytes. , 1999, The Biochemical journal.

[23]  L. Bertocci,et al.  Incorporation and utilization of [3-13C]lactate and [1,2-13C]acetate by rat skeletal muscle. , 1999, Journal of applied physiology.

[24]  G. Brooks,et al.  Role of mitochondrial lactate dehydrogenase and lactate oxidation in the intracellular lactate shuttle. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Vera,et al.  Human Monocarboxylate Transporter 2 (MCT2) Is a High Affinity Pyruvate Transporter* , 1998, The Journal of Biological Chemistry.

[26]  N. Philp,et al.  Genomic structure and developmental expression of the chicken nonocarboxylate transporter MCT3 gene. , 1998, Experimental eye research.

[27]  M. Brown,et al.  cDNA Cloning of MCT2, a Second Monocarboxylate Transporter Expressed in Different Cells than MCT1 (*) , 1995, The Journal of Biological Chemistry.

[28]  Richard G. W. Anderson,et al.  Molecular characterization of a membrane transporter for lactate, pyruvate, and other monocarboxylates: Implications for the Cori cycle , 1994, Cell.

[29]  R S Balaban,et al.  Pyruvate and lactate metabolism in the in vivo dog heart. , 1993, The American journal of physiology.

[30]  J T Reeves,et al.  Decreased reliance on lactate during exercise after acclimatization to 4,300 m. , 1991, Journal of applied physiology.

[31]  G. Brooks,et al.  Lactate and pyruvate transport is dominated by a pH gradient-sensitive carrier in rat skeletal muscle sarcolemmal vesicles. , 1990, Archives of biochemistry and biophysics.

[32]  G. Brooks,et al.  Lactate transport is mediated by a membrane-bound carrier in rat skeletal muscle sarcolemmal vesicles. , 1990, Archives of biochemistry and biophysics.

[33]  J. Wisneski,et al.  Myocardial substrate utilization during exercise in humans. Dual carbon-labeled carbohydrate isotope experiments. , 1988, The Journal of clinical investigation.

[34]  M. Rennie,et al.  L(+)-lactate transport in perfused rat skeletal muscle: kinetic characteristics and sensitivity to pH and transport inhibitors. , 1988, Biochimica et biophysica acta.

[35]  S. Spainhour,et al.  Lactate dehydrogenase in rat mitochondria. , 1987, Archives of biochemistry and biophysics.

[36]  G. Brooks,et al.  Mitochondrial reticulum in limb skeletal muscle. , 1986, The American journal of physiology.

[37]  K. S. Rogers,et al.  Localization of L-lactate dehydrogenase in mitochondria. , 1986, Archives of biochemistry and biophysics.

[38]  P. Molé,et al.  Extra O2 consumption attributable to NADH2 during maximum lactate oxidation in the heart. , 1978, Biochemical and biophysical research communications.

[39]  H. Sharma,et al.  HISTOCHEMISTRY OF LACTIC DEHYDROGENASE IN HEART AND PECTORALIS MUSCLES OF RAT , 1971, The Journal of cell biology.

[40]  S. Pande,et al.  Preferential loss of ATP-dependent long-chain fatty acid activating enzyme in mitochondria prepared using Nagarse. , 1970, Biochimica et biophysica acta.

[41]  Masaki Kobayashi,et al.  Mechanism of L-lactic acid transport in L6 skeletal muscle cells. , 2004, Drug metabolism and pharmacokinetics.