Halestrap, A. P. & Denton, R. M. Specific inhibition of pyruvate transport in rat liver mitochondria and human erythrocytes by α-cyano-4-hydroxycinnamate. Biochem. J. 138, 313–316 (1974).
Bricker, D. K. et al. A mitochondrial pyruvate carrier required for pyruvate uptake in yeast, Drosophila, and humans. Science 337, 96–100 (2012).
Herzig, S. et al. Identification and functional expression of the mitochondrial pyruvate carrier. Science 337, 93–96 (2012).
McCommis, K. S. & Finck, B. N. Mitochondrial pyruvate transport: a historical perspective and future research directions. Biochem. J. 466, 443–454 (2015).
Yiew, N. K. H. & Finck, B. N. The mitochondrial pyruvate carrier at the crossroads of intermediary metabolism. Am. J. Physiol. Endocrinol. Metab. 323, E33–E52 (2022).
Zangari, J., Petrelli, F., Maillot, B. & Martinou, J. C. The multifaceted pyruvate metabolism: role of the mitochondrial pyruvate carrier. Biomolecules 10, 1068 (2020).
Martinez-Reyes, I. & Chandel, N. S. Mitochondrial TCA cycle metabolites control physiology and disease. Nat. Commun. 11, 102 (2020).
Tavoulari, S., Sichrovsky, M. & Kunji, E. R. S. Fifty years of the mitochondrial pyruvate carrier: New insights into its structure, function, and inhibition. Acta Physiol. 238, e14016 (2023).
Papa, S., Francavilla, A., Paradies, G. & Meduri, B. The transport of pyruvate in rat liver mitochondria. FEBS Lett. 12, 285–288 (1971).
Tavoulari, S. et al. The yeast mitochondrial pyruvate carrier is a hetero-dimer in its functional state. EMBO J. 38, e100785 (2019).
Tavoulari, S. et al. Key features of inhibitor binding to the human mitochondrial pyruvate carrier hetero-dimer. Mol. Metab. 60, 101469 (2022).
Gyimesi, G. & Hediger, M. A. Sequence features of mitochondrial transporter protein families. Biomolecules 10, 1611 (2020).
Nagampalli, R. S. K. et al. Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter. Sci. Rep. 8, 3510 (2018).
Bender, T., Pena, G. & Martinou, J. C. Regulation of mitochondrial pyruvate uptake by alternative pyruvate carrier complexes. EMBO J. 34, 911–924 (2015).
Vanderperre, B. et al. MPC1-like is a placental mammal-specific mitochondrial pyruvate carrier subunit expressed in postmeiotic male germ cells. J. Biol. Chem. 291, 16448–16461 (2016).
Hegazy, L. et al. Identification of novel mitochondrial pyruvate carrier inhibitors by homology modeling and pharmacophore-based virtual screening. Biomedicines 10, 365 (2022).
Xu, L., Phelix, C. F. & Chen, L. Y. Structural insights into the human mitochondrial pyruvate carrier complexes. J. Chem. Inf. Model. 61, 5614–5625 (2021).
Schell, J. C. et al. A role for the mitochondrial pyruvate carrier as a repressor of the Warburg effect and colon cancer cell growth. Mol. Cell 56, 400–413 (2014).
McCommis, K. S. et al. Nutritional modulation of heart failure in mitochondrial pyruvate carrier-deficient mice. Nat. Metab. 2, 1232–1247 (2020).
Fernandez-Caggiano, M. et al. Mitochondrial pyruvate carrier abundance mediates pathological cardiac hypertrophy. Nat. Metab. 2, 1223–1231 (2020).
Halestrap, A. P. The mitochondrial pyruvate carrier. Kinetics and specificity for substrates and inhibitors. Biochem. J. 148, 85–96 (1975).
Vadvalkar, S. S. et al. Decreased mitochondrial pyruvate transport activity in the diabetic heart: role of mitochondrial pyruvate carrier 2 (MPC2) acetylation. J. Biol. Chem. 292, 4423–4433 (2017).
Kamm, D. R. et al. Novel insulin sensitizer MSDC-0602K improves insulinemia and fatty liver disease in mice, alone and in combination with liraglutide. J. Biol. Chem. 296, 100807 (2021).
McCommis, K. S. et al. Targeting the mitochondrial pyruvate carrier attenuates fibrosis in a mouse model of nonalcoholic steatohepatitis. Hepatology 65, 1543–1556 (2017).
Harrison, S. A. et al. Insulin sensitizer MSDC-0602K in non-alcoholic steatohepatitis: a randomized, double-blind, placebo-controlled phase IIb study. J. Hepatol. 72, 613–626 (2020).
Ghosh, A. et al. Mitochondrial pyruvate carrier regulates autophagy, inflammation, and neurodegeneration in experimental models of Parkinson’s disease. Sci. Transl. Med. 8, 368ra174 (2016).
Quansah, E. et al. Targeting energy metabolism via the mitochondrial pyruvate carrier as a novel approach to attenuate neurodegeneration. Mol. Neurodegener. 13, 28 (2018).
Shah, R. C. et al. An evaluation of MSDC-0160, a prototype mTOT modulating insulin sensitizer, in patients with mild Alzheimer’s disease. Curr. Alzheimer Res. 11, 564–573 (2014).
Olson, K. A., Schell, J. C. & Rutter, J. Pyruvate and metabolic flexibility: illuminating a path toward selective cancer therapies. Trends Biochem. Sci. 41, 219–230 (2016).
Liu, X. et al. Development of novel mitochondrial pyruvate carrier inhibitors to treat hair loss. J. Med. Chem. 64, 2046–2063 (2021).
Divakaruni, A. S. et al. Thiazolidinediones are acute, specific inhibitors of the mitochondrial pyruvate carrier. Proc. Natl Acad. Sci. USA 110, 5422–5427 (2013).
Colca, J. R. et al. Identification of a mitochondrial target of thiazolidinedione insulin sensitizers (mTOT)–relationship to newly identified mitochondrial pyruvate carrier proteins. PLoS ONE 8, e61551 (2013).
Soccio, R. E., Chen, E. R. & Lazar, M. A. Thiazolidinediones and the promise of insulin sensitization in type 2 diabetes. Cell Metab. 20, 573–591 (2014).
Kane, S. Pioglitazone drug usage statistics, United States, 2013–2022. ClinCalc.com https://clincalc.com/DrugStats/Drugs/Pioglitazone (2024).
Colca, J. R. et al. Clinical proof-of-concept study with MSDC-0160, a prototype mTOT-modulating insulin sensitizer. Clin. Pharmacol. Ther. 93, 352–359 (2013).
McCommis, K. S. et al. Loss of mitochondrial pyruvate carrier 2 in the liver leads to defects in gluconeogenesis and compensation via pyruvate-alanine cycling. Cell Metab. 22, 682–694 (2015).
Gray, L. R. et al. Hepatic mitochondrial pyruvate carrier 1 is required for efficient regulation of gluconeogenesis and whole-body glucose homeostasis. Cell Metab. 22, 669–681 (2015).
Hildyard, J. C., Ammala, C., Dukes, I. D., Thomson, S. A. & Halestrap, A. P. Identification and characterisation of a new class of highly specific and potent inhibitors of the mitochondrial pyruvate carrier. Biochim. Biophys. Acta 1707, 221–230 (2005).
Wu, X. & Rapoport, T. A. Cryo-EM structure determination of small proteins by nanobody-binding scaffolds (Legobodies). Proc. Natl Acad. Sci. USA 118, e2115001118 (2021).
Nass, K. J. et al. The role of the N-terminal amphipathic helix in bacterial YidC: Insights from functional studies, the crystal structure and molecular dynamics simulations. Biochim. Biophys. Acta Biomembr. 1864, 183825 (2022).
Miyake, T., Hizukuri, Y. & Akiyama, Y. Involvement of a membrane-bound amphiphilic helix in substrate discrimination and binding by an Escherichia coli S2P peptidase RseP. Front. Microbiol. 11, 607381 (2020).
Nepal, B., Leveritt, J. 3rd & Lazaridis, T. Membrane curvature sensing by amphipathic helices: insights from implicit membrane modeling. Biophys. J. 114, 2128–2141 (2018).
Halestrap, A. P. Pyruvate and ketone-body transport across the mitochondrial membrane. Exchange properties, pH-dependence and mechanism of the carrier. Biochem. J. 172, 377–387 (1978).
Yamashita, Y., Vinogradova, E. V., Zhang, X., Suciu, R. M. & Cravatt, B. F. A chemical proteomic probe for the mitochondrial pyruvate carrier complex. Angew. Chem. Int. Ed. Engl. 59, 3896–3899 (2020).
Wang, N. et al. Molecular basis for inhibiting human glucose transporters by exofacial inhibitors. Nat. Commun. 13, 2632 (2022).
Kumar, H. et al. Structure of sugar-bound LacY. Proc. Natl Acad. Sci. USA 111, 1784–1788 (2014).
Xu, Y. et al. Structures of bacterial homologues of SWEET transporters in two distinct conformations. Nature 515, 448–452 (2014).
Lee, Y., Nishizawa, T., Yamashita, K., Ishitani, R. & Nureki, O. Structural basis for the facilitative diffusion mechanism by SemiSWEET transporter. Nat. Commun. 6, 6112 (2015).
Paradies, G. & Papa, S. On the kinetics and substrate specificity of the pyruvate translocator in rat liver mitochondria. Biochim. Biophys. Acta 462, 333–346 (1977).
Halestrap, A. P., Brand, M. D. & Denton, R. M. Inhibition of mitochondrial pyruvate transport by phenylpyruvate and α-ketoisocaproate. Biochim. Biophys. Acta 367, 102–108 (1974).
Coleman, J. A., Green, E. M. & Gouaux, E. X-ray structures and mechanism of the human serotonin transporter. Nature 532, 334–339 (2016).
Niu, Y. et al. Structural basis of inhibition of the human SGLT2-MAP17 glucose transporter. Nature 601, 280–284 (2022).
Fan, M., Zhang, J., Lee, C. L., Zhang, J. & Feng, L. Structure and thiazide inhibition mechanism of the human Na–Cl cotransporter. Nature 614, 788–793 (2023).
Goehring, A. et al. Screening and large-scale expression of membrane proteins in mammalian cells for structural studies. Nat. Protoc. 9, 2574–2585 (2014).
Pardon, E. et al. A general protocol for the generation of nanobodies for structural biology. Nat. Protoc. 9, 674–693 (2014).
McMahon, C. et al. Yeast surface display platform for rapid discovery of conformationally selective nanobodies. Nat. Struct. Mol. Biol. 25, 289–296 (2018).
Li, Y. & Sousa, R. Expression and purification of E. coli BirA biotin ligase for in vitro biotinylation. Protein Expr. Purif. 82, 162–167 (2012).
Kowarz, E., Loscher, D. & Marschalek, R. Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol. J. 10, 647–653 (2015).
Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat. Methods 14, 290–296 (2017).
Bepler, T. et al. Positive-unlabeled convolutional neural networks for particle picking in cryo-electron micrographs. Nat. Methods 16, 1153–1160 (2019).
Wang, N. et al. Structural basis of human monocarboxylate transporter 1 inhibition by anti-cancer drug candidates. Cell 184, 370–383 e313 (2021).
Sanchez-Garcia, R. et al. DeepEMhancer: a deep learning solution for cryo-EM volume post-processing. Commun. Biol. 4, 874 (2021).
Jumper, J. et al. Highly accurate protein structure prediction with AlphaFold. Nature 596, 583–589 (2021).
Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).
Adams, P. D. et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D Biol. Crystallogr. 66, 213–221 (2010).
Davis, I. W. et al. MolProbity: all-atom contacts and structure validation for proteins and nucleic acids. Nucleic Acids Res. 35, W375–W383 (2007).
The PyMOL Molecular Graphics System, Version 1.8 (Schrodinger, LLC, 2015).
Pettersen, E. F. et al. UCSF Chimera–a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).
Pettersen, E. F. et al. UCSF ChimeraX: structure visualization for researchers, educators, and developers. Protein Sci. 30, 70–82 (2021).
Robert, X. & Gouet, P. Deciphering key features in protein structures with the new ENDscript server. Nucleic Acids Res. 42, W320–W324 (2014).
Friesner, R. A. et al. Glide: a new approach for rapid, accurate docking and scoring. 1. Method and assessment of docking accuracy. J. Med. Chem. 47, 1739–1749 (2004).
Lu, C. et al. OPLS4: improving force field accuracy on challenging regimes of chemical space. J. Chem. Theory Comput. 17, 4291–4300 (2021).
Johnston, R. C. et al. Epik: pKa and protonation state prediction through machine learning. J. Chem. Theory Comput. 19, 2380–2388 (2023).
Olsson, M. H., Sondergaard, C. R., Rostkowski, M. & Jensen, J. H. PROPKA3: consistent treatment of internal and surface residues in empirical pKa predictions. J. Chem. Theory Comput. 7, 525–537 (2011).
