TY - JOUR
T1 - Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification
AU - Dejonghe, Wim
AU - Kuenen, Sabine
AU - Mylle, Evelien
AU - Vasileva, Mina
AU - Keech, Olivier
AU - Viotti, Corrado
AU - Swerts, Jef
AU - Fendrych, Matyáš
AU - Ortiz-Morea, Fausto Andres
AU - Mishev, Kiril
AU - Delang, Simon
AU - Scholl, Stefan
AU - Zarza, Xavier
AU - Heilmann, Mareike
AU - Kourelis, Jiorgos
AU - Kasprowicz, Jaroslaw
AU - Nguyen, Le Son Long
AU - Drozdzecki, Andrzej
AU - van Houtte, Isabelle
AU - Szatmári, Anna Mária
AU - Majda, Mateusz
AU - Baisa, Gary
AU - Bednarek, Sebastian York
AU - Robert, Stéphanie
AU - Audenaert, Dominique
AU - Testerink, Christa
AU - Munnik, Teun
AU - van Damme, Daniël
AU - Heilmann, Ingo
AU - Schumacher, Karin
AU - Winne, Johan
AU - Friml, Jií
AU - Verstreken, Patrik
AU - Russinova, Eugenia
N1 - Funding information: This work was supported by the Agency for Innovation by Science and Technology for a pre-doctoral fellowship to W.D.; the Research fund KU Leuven (GOA), a Methusalem grant of the Flemish government and VIB to S.K., J.K. and P.V.; by the Netherlands Organisation for Scientific Research (NWO) for ALW grants 846.11.002 (C.T.) and 867.15.020 (T.M.); the European Research Council (project ERC-2011-StG-20101109 PSDP) (to J.F.); a European Research Council (ERC) Starting Grant (grant 260678) (to P.V.), the Research Foundation-Flanders (grants G.0747.09, G094011 and G095511) (to P.V.), the Hercules Foundation, an Interuniversity Attraction Poles Poles Program, initiated by the Belgian State, Science Policy Office (to P.V.), the Swedish VetenskapsRådet grant to O.K., the Ghent University ‘Bijzonder Onderzoek Fonds’ (BOF) for a predoctoral fellowship to F.A.O.-M., the Research Foundation-Flanders (FWO) to K.M. and E.R.
PY - 2016/6/8
Y1 - 2016/6/8
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84976491696&partnerID=8YFLogxK
U2 - 10.1038/ncomms11710
DO - 10.1038/ncomms11710
M3 - Article
C2 - 27271794
AN - SCOPUS:84976491696
SN - 2041-1723
VL - 7
JO - Nature Communications
JF - Nature Communications
M1 - 11710
ER -