[논문]Cryo-EM and MD infer water-mediated proton transport and autoinhibition mechanisms of V o complex

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

doi:10.1126/sciadv.abb9605

[해외논문] Cryo-EM and MD infer water-mediated proton transport and autoinhibition mechanisms of V o complex 원문보기

Science advances, v.6 no.41, 2020년, pp.eabb9605 -

Roh, Soung-Hun (School of Biological Sciences, Seoul National University, Seoul 08826, South Korea.) , Shekhar, Mrinal (Biodesign Institute, School of Molecular Sciences, Arizona State University, Tempe, AZ 85801, USA.) , Pintilie, Grigore (Department of Bioengineering, James H. Clark Center, Stanford University, Stanford, CA 94305, USA.) , Chipot, Christophe (Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.) , Wilkens, Stephan (Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, Syracuse, NY 13210, USA.) , Singharoy, Abhishek (Biodesign Institute, School of Molecular Sciences, Arizona State University, Tempe, AZ 85801, USA.) , Chiu, Wah (Department of Bioengineering, James H. Clark Center, Stanford University, Stanford, CA 94305, USA.)

Abstract ▼ AI-Helper

A 2.7-Å cryo-EM structure and MD simulations explain water-mediated H+ transport and autoinhibition of yeast Vo proton channel.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.

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