National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01GM058600
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
P41GM103832
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01GM079429
米国
National Institutes of Health/National Center for Research Resources (NIH/NCRR)
S10OD021600
米国
National Research Foundation (NRF, Korea)
NRF-2019M3E5D6063865
韓国
National Science Foundation (NSF, United States)
MCB-1942763
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01GM067887
米国
Agence Nationale de la Recherche (ANR)
ProteaseInAction Grant
フランス
引用
ジャーナル: Sci Adv / 年: 2020 タイトル: Cryo-EM and MD infer water-mediated proton transport and autoinhibition mechanisms of V complex. 著者: Soung-Hun Roh / Mrinal Shekhar / Grigore Pintilie / Christophe Chipot / Stephan Wilkens / Abhishek Singharoy / Wah Chiu / 要旨: Rotary vacuolar adenosine triphosphatases (V-ATPases) drive transmembrane proton transport through a V proton channel subcomplex. Despite recent high-resolution structures of several rotary ATPases, ...Rotary vacuolar adenosine triphosphatases (V-ATPases) drive transmembrane proton transport through a V 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 V 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 V that shows the -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 -ring rotation. Our cryo-EM models and simulations also rationalize a mechanism for inhibition of passive proton transport as observed for free V that is generated as a result of V-ATPase regulation by reversible disassembly in vivo.