Structure of the Lipid Nanodisc-reconstituted Vacuolar ATPase Proton Channel: DEFINITION OF THE INTERACTION OF ROTOR AND STATOR AND IMPLICATIONS FOR ENZYME REGULATION BY REVERSIBLE DISSOCIATION.

ABSTRACT: Eukaryotic vacuolar H⁺-ATPase (V-ATPase) is a multisubunit enzyme complex that acidifies subcellular organelles and the extracellular space. V-ATPase consists of soluble V₁-ATPase and membrane-integral V_o proton channel sectors. To investigate the mechanism of V-ATPase regulation by reversible disassembly, we recently determined a cryo-EM reconstruction of yeast V_o The structure indicated that, when V₁ is released from V_o, the N-terminal cytoplasmic domain of subunit a (a_NT) changes conformation to bind rotor subunit d However, insufficient resolution precluded a precise definition of the a_NT-d interface. Here we reconstituted V_o into lipid nanodiscs for single-particle EM. 3D reconstructions

SUBMITTER: Stam NJ 

PROVIDER: S-EPMC5290949 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature