regulation of the v-atpase along the endocytic pathway occurs through reversible subunit association and membrane localization监管v-atpase沿着内吞作用的途径通过发生可逆的亚基协会和膜定位.pdfVIP

Regulation of the V-ATPase along the Endocytic Pathway Occurs through Reversible Subunit Association and Membrane Localization ´ 1 1 2 ´ 2 1 Celine Lafourcade , Komla Sobo , Sylvie Kieffer-Jaquinod , Jerome Garin , F. Gisou van der Goot * ´ ´ 1 Global Health Institute, Ecole Polytechnique Federale de Lausanne, Faculty of Life Sciences, Lausanne, Switzerland, 2 CEA, DSV, iRTSV, Laboratoire d’Etude de la ´ Dynamique des Proteomes, INSERM U880, Grenoble, France Abstract The lumen of endosomal organelles becomes increasingly acidic when going from the cell surface to lysosomes. Luminal pH thereby regulates important processes such as the release of internalized ligands from their receptor or the activation of lysosomal enzymes. The main player in endosomal acidification is the vacuolar ATPase (V-ATPase), a multi-subunit transmembrane complex that pumps protons from the cytoplasm to the lumen of organelles, or to the outside of the cell. The active V-ATPase is composed of two multi-subunit domains, the transmembrane V and the cytoplasmic V . Here we 0 1 found that the ratio of membrane associated V1/Vo varies along the endocytic pathway, the relative abundance of V1 being higher on late endosomes than on early endosomes, providing an explanation for the higher acidity of late endosomes. We also found that all membrane-bound V-ATPase subunits were associated with detergent resistant membranes (DRM) isolated from late endosomes,