Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification

article Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification published yes Wim Dejonghe author Sabine Kuenen author Evelien Mylle author Mina K Vasileva author 3407EB18-F248-11E8-B48F-1D18A9856A87 Olivier Keech author Corrado Viotti author Jef Swerts author Matyas Fendrych author 43905548-F248-11E8-B48F-1D18A9856A870000-0002-9767-8699 Fausto Ortiz Morea author Kiril Mishev author Simon Delang author Stefan Scholl author Xavier Zarza author Mareike Heilmann author Jiorgos Kourelis author Jaroslaw Kasprowicz author Le Nguyen author Andrzej Drozdzecki author Isabelle Van Houtte author Anna Szatmári author Mateusz Majda author Gary Baisa author Sebastian Bednarek author Stéphanie Robert author Dominique Audenaert author Christa Testerink author Teun Munnik author Daniël Van Damme author Ingo Heilmann author Karin Schumacher author Johan Winne author Jirí Friml author 4159519E-F248-11E8-B48F-1D18A9856A870000-0002-8302-7596 Patrik Verstreken author Eugenia Russinova author JiFr department Polarity and subcellular dynamics in plants project ATP production requires the establishment of an electrochemical proton gradient across the inner mitochondrial membrane. Mitochondrial uncouplers dissipate this proton gradient and disrupt numerous cellular processes, including vesicular trafficking, mainly through energy depletion. Here we show that Endosidin9 (ES9), a novel mitochondrial uncoupler, is a potent inhibitor of clathrin-mediated endocytosis (CME) in different systems and that ES9 induces inhibition of CME not because of its effect on cellular ATP, but rather due to its protonophore activity that leads to cytoplasm acidification. We show that the known tyrosine kinase inhibitor tyrphostinA23, which is routinely used to block CME, displays similar properties, thus questioning its use as a specific inhibitor of cargo recognition by the AP-2 adaptor complex via tyrosine motif-based endocytosis signals. Furthermore, we show that cytoplasm acidification dramatically affects the dynamics and recruitment of clathrin and associated adaptors, and leads to reduction of phosphatidylinositol 4,5-biphosphate from the plasma membrane. https://research-explorer.ista.ac.at/download/1346/5369/IST-2016-653-v1+1_ncomms11710_1_.pdf application/pdfno https://creativecommons.org/licenses/by/4.0/ Nature Publishing Group2016 eng Nature Communications 00037789980000110.1038/ncomms11710 7 https://research-explorer.ista.ac.at/record/7172 Dejonghe W, Kuenen S, Mylle E, Vasileva MK, Keech O, Viotti C, Swerts J, Fendrych M, Ortiz Morea F, Mishev K, Delang S, Scholl S, Zarza X, Heilmann M, Kourelis J, Kasprowicz J, Nguyen L, Drozdzecki A, Van Houtte I, Szatmári A, Majda M, Baisa G, Bednarek S, Robert S, Audenaert D, Testerink C, Munnik T, Van Damme D, Heilmann I, Schumacher K, Winne J, Friml J, Verstreken P, Russinova E. 2016. Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification. Nature Communications. 7, 11710. Dejonghe W, Kuenen S, Mylle E, et al. Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification. Nature Communications. 2016;7. doi:<a href="https://doi.org/10.1038/ncomms11710">10.1038/ncomms11710</a> Dejonghe, Wim, Sabine Kuenen, Evelien Mylle, Mina K Vasileva, Olivier Keech, Corrado Viotti, Jef Swerts, et al. “Mitochondrial Uncouplers Inhibit Clathrin-Mediated Endocytosis Largely through Cytoplasmic Acidification.” Nature Communications. Nature Publishing Group, 2016. <a href="https://doi.org/10.1038/ncomms11710">https://doi.org/10.1038/ncomms11710</a>. Dejonghe, Wim, et al. “Mitochondrial Uncouplers Inhibit Clathrin-Mediated Endocytosis Largely through Cytoplasmic Acidification.” Nature Communications, vol. 7, 11710, Nature Publishing Group, 2016, doi:<a href="https://doi.org/10.1038/ncomms11710">10.1038/ncomms11710</a>. Dejonghe, W., Kuenen, S., Mylle, E., Vasileva, M. K., Keech, O., Viotti, C., … Russinova, E. (2016). Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification. Nature Communications. Nature Publishing Group. <a href="https://doi.org/10.1038/ncomms11710">https://doi.org/10.1038/ncomms11710</a> W. Dejonghe, S. Kuenen, E. Mylle, M.K. Vasileva, O. Keech, C. Viotti, J. Swerts, M. Fendrych, F. Ortiz Morea, K. Mishev, S. Delang, S. Scholl, X. Zarza, M. Heilmann, J. Kourelis, J. Kasprowicz, L. Nguyen, A. Drozdzecki, I. Van Houtte, A. Szatmári, M. Majda, G. Baisa, S. Bednarek, S. Robert, D. Audenaert, C. Testerink, T. Munnik, D. Van Damme, I. Heilmann, K. Schumacher, J. Winne, J. Friml, P. Verstreken, E. Russinova, Nature Communications 7 (2016). W. Dejonghe et al., “Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification,” Nature Communications, vol. 7. Nature Publishing Group, 2016. 13462018-12-11T11:51:30Z2026-04-08T13:54:44Z