
(C, D) Lipid molecules inside the c-ring at the luminal leaflet (C) and (B) Electrostatic surface representation of subunits a, e, RNAseK, andĬ-ring, showing the interactions between the lipids and protein subunits. PC: phosphatidylcholine PE: phosphatidylethanolamine PS: These lipids establish extensive interactions with the protein subunitsĪbove. (A) Lipid molecules at the interfaces of subunits a, e, RNAseK, and theĬ-ring. Subunit (Y313 and E346) and with neighboring subunits. (J) Detailed interactions of the mutation sites within the ATP6AP1 (I) Missense disease mutations of ATP6AP1 (red) mapped onto its ATP6AP1 is the mostĬonnected subunit in the diagram. Subunits, ATP6AP1, ATP6AP2, subunit d, and lipids. (H) A schematic diagram of the interaction network involving c-ring (G) Surface representation of the c-ring, ATP6AP1, ATP6AP2, and subunitĭ, showing their interactions. (green), showing the overall interactions of ATP6AP1 with neighboring (E, F) Side view (E) and cytosolic view (F) of the c-ring (c” inīlue and c subunits in slate), ATP6AP1 (yellow), ATP6AP2 (orange), and subunit d LAMP-1 (green) obtained from the DALI server (Holm and Sander, 1995). (D) Structural alignment of ATP6AP1 luminal domain (LD, yellow) with Location of aĬonserved disulfide bond between C371 and C418 is marked. (C) Ribbon diagram of the luminal domain of ATP6AP1. (B) Ribbon diagram of mature ATP6AP1 in the human V-ATPase TheĬleavage sites for furin or another protease in their luminal domains are (A) Domain diagrams of human ATP6AP1 and ATP6AP2, and yeast ATP6AP1.

This study identifies mechanisms of V-ATPase assembly and biogenesis that rely on the integrated roles of ATP6AP1, glycans, and lipids.Ĭopyright © 2020 Elsevier Inc. The glycolipids and the glycosylated V o subunits form a luminal glycan coat critical for V-ATPase folding, localization, and stability. We define ATP6AP1 as a structural hub for V o complex assembly because it connects to multiple V o subunits and phospholipids in the c-ring. Aided by mass spectrometry, we build all known protein subunits with associated N-linked glycans and identify glycolipids and phospholipids in the V o complex. Here, we report cryoelectron microscopy structures of human V-ATPase in three rotational states at up to 2.9-Å resolution. They play important roles in acidification of intracellular vesicles, organelles, and the extracellular milieu in eukaryotes. Electronic address: or vacuolar-type adenosine triphosphatases (V-ATPases) are ATP-driven proton pumps comprised of a cytoplasmic V 1 complex for ATP hydrolysis and a membrane-embedded V o complex for proton transfer. Electronic address: 4 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH 43210, USA The Ohio State University Comprehensive Cancer Center, Columbus, OH 43210, USA.

Electronic address: 2 Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3QZ, UK. 1 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA.
