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Mol Cell. 2017 Aug 17;67(4):711-723.e7. doi: 10.1016/j.molcel.2017.07.019.

Systematic Identification of MCU Modulators by Orthogonal Interspecies Chemical Screening.

Author information

1
Gene Center/Department of Biochemistry, Ludwig-Maximilians Universität München, Munich 81377, Germany; Institute for Diabetes and Obesity, Helmholtz Zentrum München, Neuherberg 85764, Germany.
2
Department of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
3
Instituto de Biología y Genética Molecular (IBGM), University of Valladolid and Consejo Superior de Investigaciones Científicas (CSIC), Valladolid 47003, Spain.
4
Chair for Proteomics and Bioanalytics, Technical University Munich, Freising 85354, Germany.
5
Chair for Proteomics and Bioanalytics, Technical University Munich, Freising 85354, Germany; Center for Integrated Protein Science Munich, Freising 85354, Germany.
6
Biomedical Center, Department of Physiological Chemistry, Ludwig-Maximilians Universität München, Martinsried 81377, Germany.
7
Department of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
8
Gene Center/Department of Biochemistry, Ludwig-Maximilians Universität München, Munich 81377, Germany; Institute for Diabetes and Obesity, Helmholtz Zentrum München, Neuherberg 85764, Germany. Electronic address: perocchi@genzentrum.lmu.de.

Abstract

The mitochondrial calcium uniporter complex is essential for calcium (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 for targeting uniporter activity. Here we introduce a high-throughput assay that selects for human MCU-specific small-molecule modulators in primary drug screens. Using isolated yeast mitochondria, reconstituted with human MCU, its essential regulator EMRE, and aequorin, and exploiting a D-lactate- and mannitol/sucrose-based bioenergetic shunt that greatly minimizes false-positive hits, we identify mitoxantrone out of more than 600 clinically approved drugs as a direct selective inhibitor of human MCU. We validate mitoxantrone in orthogonal mammalian cell-based assays, demonstrating that our screening approach is an effective and robust tool for MCU-specific drug discovery and, more generally, for the identification of compounds that target mitochondrial functions.

KEYWORDS:

MCU; bioenergetics; calcium; calcium signaling; drug discovery; drug screening; high-throughput screening; mitochondria; mitochondrial calcium uniporter

PMID:
28820965
PMCID:
PMC5825229
DOI:
10.1016/j.molcel.2017.07.019
[Indexed for MEDLINE]
Free PMC Article

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