Cryo-EM and MD infer water-mediated proton transport and autoinhibition mechanisms of Vo complex

Soung Hun Roh, Mrinal Shekhar, Grigore Pintilie, Christophe Chipot, Stephan Wilkens, Abhishek Singharoy, Wah Chiu

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Rotary vacuolar adenosine triphosphatases (V-ATPases) drive transmembrane proton transport through a Vo proton channel subcomplex. Despite recent high-resolution structures of several rotary ATPases, the dynamic mechanism of proton pumping remains elusive. Here, we determined a 2.7-Å cryo-electron microscopy (cryo-EM) structure of yeast Vo proton channel in nanodisc that reveals the location of ordered water molecules along the proton path, details of specific protein-lipid interactions, and the architecture of the membrane scaffold protein. Moreover, we uncover a state of Vo that shows the c-ring rotated by ∼14°. Molecular dynamics simulations demonstrate that the two rotary states are in thermal equilibrium and depict how the protonation state of essential glutamic acid residues couples water-mediated proton transfer with c-ring rotation. Our cryo-EM models and simulations also rationalize a mechanism for inhibition of passive proton transport as observed for free Vo that is generated as a result of V-ATPase regulation by reversible disassembly in vivo.

Original languageEnglish (US)
Article numbereabb9605
JournalScience Advances
Volume6
Issue number41
DOIs
StatePublished - Oct 7 2020

ASJC Scopus subject areas

  • General

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