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 language | English (US) |
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Article number | eabb9605 |
Journal | Science Advances |
Volume | 6 |
Issue number | 41 |
DOIs | |
State | Published - Oct 7 2020 |
ASJC Scopus subject areas
- General
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Dive into the research topics of 'Cryo-EM and MD infer water-mediated proton transport and autoinhibition mechanisms of Vo complex'. Together they form a unique fingerprint.Datasets
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3.6A Yeast Vo state3 prime
Roh, S. (Contributor), Shekhar, M. (Contributor), Pintilie, G. (Contributor), Chipot, C. (Contributor), Wilkens, S. (Contributor), Singharoy, A. (Contributor) & Chiu, W. (Contributor), Protein Data Bank (PDB), Nov 4 2020
DOI: 10.2210/pdb6M0S, https://www.wwpdb.org/pdb?id=pdb_00006m0s
Dataset
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2.7A Yeast Vo state3
Roh, S. (Contributor), Shekhar, M. (Contributor), Pintilie, G. (Contributor), Chipot, C. (Contributor), Wilkens, S. (Contributor), Singharoy, A. (Contributor) & Chiu, W. (Contributor), Protein Data Bank (PDB), Nov 4 2020
DOI: 10.2210/pdb6M0R, https://www.wwpdb.org/pdb?id=pdb_00006m0r
Dataset