Embryonic Lethality of Mitochondrial Pyruvate Carrier 1 Deficient Mouse Can Be Rescued by a Ketogenic Diet

Vanderperre, Benoît; Herzig, Sébastien; Krznar, Petra; Hörl, Manuel; Ammar, Zeinab; Montessuit, Sylvie; Pierredon, Sandra; Zamboni, Nicola; Martinou, Jean‐Claude

doi:10.1371/journal.pgen.1006056

Cited by 60 publications

(74 citation statements)

References 47 publications

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“…Soluble and integral membrane proteins were separated by ultracentrifugation at 100,000 ϫ g for 30 min. Cytosolic proteins and soluble mitochondrial proteins were submitted to chloroform-methanol precipitation before SDS-PAGE and Western blotting as described elsewhere (25). The proteinase K accessibility test was performed as described previously (46).…”

Section: Methodsmentioning

confidence: 99%

“…To confirm that MPC1L can sustain pyruvate import into mammalian mitochondria, we took advantage of previously described MPC-deficient mouse embryonic fibroblasts (MEFs) in which MPC1 expression had been ablated using a gene trap strategy (17,25). These cells display no detectable MPC1 by Western blotting analysis (Fig.…”

Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tmentioning

confidence: 99%

“…3D). Furthermore, the MPC2 protein was also undetectable, despite the presence of its mRNA (25), presumably because MPC2 is degraded in the absence of MPC1. Interestingly, Western blots using our inhouse antibody against the C terminus of mouse MPC1L, showed no compensatory expression of the endogenous MPC1L protein in the mitochondria-rich fraction from these cells (Fig.…”

Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tmentioning

confidence: 99%

“…Generation of immortalized MPC1 KO MEFs was described previously (25). HEK293T, HeLa, and MEFs cell lines were cultured in DMEM containing 4.5 g/liter glucose, 2 mM glutamine, 10% FBS, and 1% penicillin/streptomycin (Invitrogen).…”

Section: Preparation Of Lysates From Mouse Tissues and Spermmentioning

confidence: 99%

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MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Vanderperre¹,

Cermakova²,

Escoffier

et al. 2016

Journal of Biological Chemistry

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Selective transport of pyruvate across the inner mitochondrial membrane by the mitochondrial pyruvate carrier (MPC) is a fundamental step that couples cytosolic and mitochondrial metabolism. The recent molecular identification of the MPC complex has revealed two interacting subunits, MPC1 and MPC2. Although in yeast, an additional subunit, MPC3, can functionally replace MPC2, no alternative MPC subunits have been described in higher eukaryotes. Here, we report for the first time the existence of a novel MPC subunit termed MPC1-like (MPC1L), which is present uniquely in placental mammals. MPC1L shares high sequence, structural, and topological homology with MPC1. In addition, we provide several lines of evidence to show that MPC1L is functionally equivalent to MPC1: 1) when co-expressed with MPC2, it rescues pyruvate import in a MPC-deleted yeast strain; 2) in mammalian cells, it can associate with MPC2 to form a functional carrier as assessed by bioluminescence resonance energy transfer; 3) in MPC1 depleted mouse embryonic fibroblasts, MPC1L rescues the loss of pyruvate-driven respiration and stabilizes MPC2 expression; and 4) MPC1-and MPC1L-mediated pyruvate imports show similar efficiency. However, we show that MPC1L has a highly specific expression pattern and is localized almost exclusively in testis and more specifically in postmeiotic spermatids and sperm cells. This is in marked contrast to MPC1/MPC2, which are ubiquitously expressed throughout the organism. To date, the biological importance of this alternative MPC complex during spermatogenesis in placental mammals remains unknown. Nevertheless, these findings open up new avenues for investigating the structure-function relationship within the MPC complex.The metabolic balance between glycolysis and oxidative phosphorylation (OXPHOS) 2 is of crucial importance in determining cell function and fate. Rapidly proliferating cells rely preferentially on glycolysis despite its lower yield of ATP, because glycolytic intermediates efficiently meet the anabolic needs for sustained cell growth. In rapidly dividing cells, this occurs even in the presence of oxygen, a process called aerobic glycolysis, or the Warburg effect (1, 2). Differentiated cells rely more heavily on OXPHOS to meet the energy demands associated with their specialization. In addition, a shift from glycolytic to oxidative metabolism is necessary for the differentiation process to occur (2-4), and conversely, a hallmark of cancer lies in the rewiring of cellular metabolism toward increased glycolytic flux (5). Pyruvate, the end product of glycolysis, is imported into the mitochondrial matrix where it fuels the TCA cycle and thereby provides electrons and reducing equivalents to the respiratory chain. Pyruvate-derived acetyl-CoA can also be used as an anabolic substrate for the synthesis of lipids and amino acids. Thus, elucidating the molecular mechanisms that determine the intracellular fate of pyruvate is important for understanding the control of cell metabolism and ultimately of cell funct...

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Section: Methodsmentioning

confidence: 99%

Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tmentioning

confidence: 99%

Section: A a A T Ag L Tq R -------------------P Yg C --------A T T Tmentioning

confidence: 99%

Section: Preparation Of Lysates From Mouse Tissues and Spermmentioning

confidence: 99%

See 2 more Smart Citations

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Vanderperre¹,

Cermakova²,

Escoffier

et al. 2016

Journal of Biological Chemistry

Self Cite

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show abstract

“…ATP levels were assessed in INS-1E cells using a Seahorse XF e 24 Flux Analyzer (Seahorse Biosciences) as described (Vanderperre et al, 2016). Amino acid measurements were done by reverse-phase highperformance liquid chromatography (HPLC) after derivatization with O-phthalaldehyde (Vetterli et al, 2012).…”

Section: Insulin Secretion Calcium Measurements Atp and Amino Acidmentioning

confidence: 99%

Epigallocatechin-3-gallate (EGCG) activates AMPK through the inhibition of glutamate dehydrogenase in muscle and pancreatic ß-cells: A potential beneficial effect in the pre-diabetic state?

Pournourmohammadi

Grimaldi

Stridh

et al. 2017

The International Journal of Biochemistry & Cell Biology

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a b s t r a c tGlucose homeostasis is determined by insulin secretion from the ß-cells in pancreatic islets and by glucose uptake in skeletal muscle and other insulin target tissues. While glutamate dehydrogenase (GDH) senses mitochondrial energy supply and regulates insulin secretion, its role in the muscle has not been elucidated. Here we investigated the possible interplay between GDH and the cytosolic energy sensing enzyme 5 -AMP kinase (AMPK), in both isolated islets and myotubes from mice and humans. The green tea polyphenol epigallocatechin-3-gallate (EGCG) was used to inhibit GDH. Insulin secretion was reduced by EGCG upon glucose stimulation and blocked in response to glutamine combined with the allosteric GDH activator BCH (2-aminobicyclo-[2,2,1] heptane-2-carboxylic acid). Insulin secretion was similarly decreased in islets of mice with ß-cell-targeted deletion of GDH (ßGlud1 −/− ). EGCG did not further reduce insulin secretion in the mutant islets, validating its specificity. In human islets, EGCG attenuated both basal and nutrient-stimulated insulin secretion. Glutamine/BCH-induced lowering of AMPK phosphorylation did not operate in ßGlud1 −/− islets and was similarly prevented by EGCG in control islets, while high glucose systematically inactivated AMPK. In mouse C2C12 myotubes, like in islets, the inhibition of AMPK following GDH activation with glutamine/BCH was reversed by EGCG. Stimulation of GDH in primary human myotubes caused lowering of insulin-induced 2-deoxy-glucose uptake, partially counteracted by EGCG. Thus, mitochondrial energy provision through anaplerotic input via GDH influences the activity of the cytosolic energy sensor AMPK. EGCG may be useful in obesity by resensitizing insulin-resistant muscle while blunting hypersecretion of insulin in hypermetabolic states.

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Identification and characterization of novel MPC1 gene variants causing mitochondrial pyruvate carrier deficiency

Jiang

Alahmad

et al. 2022

J of Inher Metab Disea

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Pyruvate, the end product of glycolysis, is a key metabolic molecule enabling mitochondrial adenosine triphosphate synthesis and takes part in multiple biosynthetic pathways within mitochondria. The mitochondrial pyruvate carrier (MPC) plays a vital role in transporting pyruvate from the cytosol into the organelle. In humans, MPC is a hetero-oligomeric complex formed by the MPC1 and MPC2 paralogs that are both necessary to stabilize each other and form a functional MPC. MPC deficiency (OMIM#614741) due to pathogenic MPC1 variants is a rare autosomal recessive disease involving developmental delay, microcephaly, growth failure, and increased serum lactate and pyruvate. To date, two MPC1 variants in four cases have been reported, though only one with a detailed clinical description. Herein, we report three novel pathogenic MPC1 variants in six patients from three unrelated families, identified within European, Kuwaiti, and Chinese mitochondrial disease patient cohorts, one of whom presented as a Leigh-like syndrome. Functional analysis in primary fibroblasts from the patients revealed decreased expression of MPC1 and MPC2. We rescued pyruvate-driven oxygen consumption rate in patient's fibroblasts by reconstituting with wild-type MPC1. Complementing homozygous MPC1 mutant cDNA with CRISPR-deleted MPC1 C2C12 cells verified the

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Embryonic Lethality of Mitochondrial Pyruvate Carrier 1 Deficient Mouse Can Be Rescued by a Ketogenic Diet

Cited by 60 publications

References 47 publications

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Epigallocatechin-3-gallate (EGCG) activates AMPK through the inhibition of glutamate dehydrogenase in muscle and pancreatic ß-cells: A potential beneficial effect in the pre-diabetic state?

Identification and characterization of novel MPC1 gene variants causing mitochondrial pyruvate carrier deficiency

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