Abstract
The metabolic cofactor coenzyme A (CoA) gained renewed attention because of its roles in neurodegeneration, protein acetylation, autophagy and signal transduction. The long-standing dogma is that eukaryotic cells obtain CoA exclusively via the uptake of extracellular precursors, especially vitamin B5, which is intracellularly converted through five conserved enzymatic reactions into CoA. This study demonstrates an alternative mechanism that allows cells and organisms to adjust intracellular CoA levels by using exogenous CoA. Here CoA was hydrolyzed extracellularly by ectonucleotide pyrophosphatases to 4′-phosphopantetheine, a biologically stable molecule able to translocate through membranes via passive diffusion. Inside the cell, 4′-phosphopantetheine was enzymatically converted back to CoA by the bifunctional enzyme CoA synthase. Phenotypes induced by intracellular CoA deprivation were reversed when exogenous CoA was provided. Our findings answer long-standing questions in fundamental cell biology and have major implications for the understanding of CoA-related diseases and therapies.
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Acknowledgements
This work was supported by the European Commission's Seventh Framework Programme (grant number FP7/2007-2013, HEALTH-F2-2011, grant agreement number 277984, TIRCON to V.T., H.P., S.H. and O.C.M.S.), a GUIDE research school grant (to B.S.), a VICI grant (NWO-grant 865.10.012 to O.S.) and Telethon GGP11088 (to V.T.). Part of the work was performed at the UMCG Microscopy and Imaging Center (UMIC), which is sponsored by NWO grant 175-010-2009-023. We thank P.G. Tepper and R. Van Merkerk (Pharmaceutical Biology, University of Groningen) for providing HPLC technical assistance and J.-W. Kok and D. Hoekstra (Department of Cell Biology, UMCG) for helpful discussions. We also thank T. de Boer and F. Oostebring from the Analytical Biochemical Laboratory BV (ABL, Assen, the Netherlands) for mass spectrometry analysis. We are grateful to the PKAN patients and their families who contributed samples to this study.
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B.S., M.B., R.A.L., G.K., H.P., S.H., V.T. and O.C.M.S. designed the research. B.S., M.B., M.v.d.Z., B.K., C.C., R.A.L., O.S., A.P. and N.A.G. performed experiments. B.S., M.B., R.A.L. and O.C.M.S. analyzed results. E.A.A.N., G.K., H.P., S.H., V.T., D.-J.R., N.A.G. and O.C.M.S. supervised the research. B.S., N.A.G. and O.C.M.S. wrote the manuscript.
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B.S., O.C.M.S., G.K., H.P. and A.P. are co-inventors on European patent application EP 2 868 662 A1. B.S., O.C.M.S., G.K. and H.P. are co-inventors on Slovenian patent application P-201400452. B.S., O.C.M.S., G.K., H.P. and S.H. are co-inventors on European patent application EP 15468006.0. H.P. and G.K. are shareholders in Acies Bio, Ltd.
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Srinivasan, B., Baratashvili, M., van der Zwaag, M. et al. Extracellular 4′-phosphopantetheine is a source for intracellular coenzyme A synthesis. Nat Chem Biol 11, 784–792 (2015). https://doi.org/10.1038/nchembio.1906
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DOI: https://doi.org/10.1038/nchembio.1906
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