Systematic Identification of MCU Modulators by Orthogonal Interspecies Chemical Screening

摘要: The mitochondrial calcium uniporter complex is essential for (Ca2+) uptake into mitochondria of all mammalian tissues, where it regulates bioenergetics, cell death, and Ca2+ signal transduction. Despite its involvement in several human diseases, we currently lack pharmacological agents targeting activity. Here introduce a high-throughput assay that selects MCU-specific small-molecule modulators primary drug screens. Using isolated yeast mitochondria, reconstituted with MCU, regulator EMRE, aequorin, exploiting D-lactate- mannitol/sucrose-based bioenergetic shunt greatly minimizes false-positive hits, identify mitoxantrone out more than 600 clinically approved drugs as direct selective inhibitor MCU. We validate mitoxantrone orthogonal cell-based assays, demonstrating our screening approach an effective robust tool MCU-specific discovery and, generally, the identification compounds target functions.