Transport of pyruvate into mitochondria by the mitochondrial pyruvate carrier is crucial for complete oxidation of glucose and for biosynthesis of amino acids and lipids. Zaprinast is a well known phosphodiesterase inhibitor and lead compound for sildenafil. We found Zaprinast alters the metabolomic profile of mitochondrial intermediates and amino acids in retina and brain. This metabolic effect of Zaprinast does not depend on inhibition of phosphodiesterase activity. By providing 13C-labeled glucose and glutamine as fuels, we found that the metabolic profile of the Zaprinast effect is nearly identical to that of inhibitors of the mitochondrial pyruvate carrier. Both stimulate oxidation of glutamate and massive accumulation of aspartate. Moreover, Zaprinast inhibits pyruvate-driven O2 consumption in brain mitochondria and blocks mitochondrial pyruvate carrier in liver mitochondria. Inactivation of the aspartate glutamate carrier in retina does not attenuate the metabolic effect of Zaprinast. Our results show that Zaprinast is a potent inhibitor of mitochondrial pyruvate carrier activity, and this action causes aspartate to accumulate at the expense of glutamate. Our findings show that Zaprinast is a specific mitochondrial pyruvate carrier (MPC) inhibitor and may help to elucidate the roles of MPC in amino acid metabolism and hypoglycemia.
This work was supported, in whole or in part, by National Institutes of Health Grants EY06641 and EY017863 (to J. B. H.). This work was also supported by grants from the Deutsche Forschungsgemeinschaft (Pa1751/4-1), EU (DRUGSFORD: HEALTH-F2-2012-304963), and the Kerstan Foundation (to F. P.-D.) and the Ministerio de Economíay Competitividad (BFU2011-30456-C02-01/BMC) and Coumunidad Autónoma de Madrid (S2010/BMD-2402) (to J. S.). This work was also funded by the CIBERER, an initiative from the Instituto de Salud Carlos III, and an institutional grant from the Fundación Ramón Areces to the Centro de Biología Molecular Severo Ochoa.
