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Title: The 3.5-Å CryoEM Structure of Nanodisc-Reconstituted Yeast Vacuolar ATPase Vo Proton Channel

Abstract

The molecular mechanism of transmembrane proton translocation in rotary motor ATPases is not fully understood. Here in this paper, we report the 3.5-Å resolution cryoEM structure of the lipid nanodisc-reconstituted Vo proton channel of the yeast vacuolar H+-ATPase, captured in a physiologically relevant, autoinhibited state. The resulting atomic model provides structural detail for the amino acids that constitute the proton pathway at the interface of the proteolipid ring and subunit a. Based on the structure and previous mutagenesis studies, we propose the chemical basis of transmembrane proton transport. Moreover, we discovered that the C terminus of the assembly factor Voa1 is an integral component of mature Vo. Voa1’s C-terminal transmembrane α helix is bound inside the proteolipid ring, where it contributes to the stability of the complex. Our structure rationalizes possible mechanisms by which mutations in human Vo can result in disease phenotypes and may thus provide new avenues for therapeutic interventions.

Authors:
 [1];  [2];  [3];  [2];  [4];  [5];  [2]
  1. Stanford Univ., CA (United States). Dept. of Bioengineering and James H. Clark Center; SLAC National Accelerator Lab., Menlo Park, CA (United States). Biosciences Division
  2. State Univ. of New York (SUNY) Upstate Medical University, Syracuse, NY (United States). Dept. of Biochemistry and Molecular Biology
  3. Baylor College of Medicine, Houston, TX (United States). Graduate Program in Quantitative and Computational Biosciences; Baylor College of Medicine, Houston, TX (United States). Verna and Marrs McLean Dept. of Biochemistry and Molecular Biology
  4. Baylor College of Medicine, Houston, TX (United States). Verna and Marrs McLean Dept. of Biochemistry and Molecular Biology
  5. Stanford Univ., CA (United States). Dept. of Bioengineering and James H. Clark Center; SLAC National Accelerator Lab., Menlo Park, CA (United States). Biosciences Division; Baylor College of Medicine, Houston, TX (United States). Verna and Marrs McLean Dept. of Biochemistry and Molecular Biology
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1475581
Grant/Contract Number:  
AC02-76SF00515; GM058600; P41GM103832; R01GM079429; T15LM007093
Resource Type:
Accepted Manuscript
Journal Name:
Molecular Cell
Additional Journal Information:
Journal Volume: 69; Journal Issue: 6; Journal ID: ISSN 1097-2765
Publisher:
Cell Press - Elsevier
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; vacuolar H+-ATPase; Vo proton channelcryo; EMlipid nanodisc; V-ATPase assembly; Voa1; membrane protein structure; proton pumping; reversible disassembly

Citation Formats

Roh, Soung-Hun, Stam, Nicholas J., Hryc, Corey F., Couoh-Cardel, Sergio, Pintilie, Grigore, Chiu, Wah, and Wilkens, Stephan. The 3.5-Å CryoEM Structure of Nanodisc-Reconstituted Yeast Vacuolar ATPase Vo Proton Channel. United States: N. p., 2018. Web. doi:10.1016/j.molcel.2018.02.006.
Roh, Soung-Hun, Stam, Nicholas J., Hryc, Corey F., Couoh-Cardel, Sergio, Pintilie, Grigore, Chiu, Wah, & Wilkens, Stephan. The 3.5-Å CryoEM Structure of Nanodisc-Reconstituted Yeast Vacuolar ATPase Vo Proton Channel. United States. doi:10.1016/j.molcel.2018.02.006.
Roh, Soung-Hun, Stam, Nicholas J., Hryc, Corey F., Couoh-Cardel, Sergio, Pintilie, Grigore, Chiu, Wah, and Wilkens, Stephan. Thu . "The 3.5-Å CryoEM Structure of Nanodisc-Reconstituted Yeast Vacuolar ATPase Vo Proton Channel". United States. doi:10.1016/j.molcel.2018.02.006. https://www.osti.gov/servlets/purl/1475581.
@article{osti_1475581,
title = {The 3.5-Å CryoEM Structure of Nanodisc-Reconstituted Yeast Vacuolar ATPase Vo Proton Channel},
author = {Roh, Soung-Hun and Stam, Nicholas J. and Hryc, Corey F. and Couoh-Cardel, Sergio and Pintilie, Grigore and Chiu, Wah and Wilkens, Stephan},
abstractNote = {The molecular mechanism of transmembrane proton translocation in rotary motor ATPases is not fully understood. Here in this paper, we report the 3.5-Å resolution cryoEM structure of the lipid nanodisc-reconstituted Vo proton channel of the yeast vacuolar H+-ATPase, captured in a physiologically relevant, autoinhibited state. The resulting atomic model provides structural detail for the amino acids that constitute the proton pathway at the interface of the proteolipid ring and subunit a. Based on the structure and previous mutagenesis studies, we propose the chemical basis of transmembrane proton transport. Moreover, we discovered that the C terminus of the assembly factor Voa1 is an integral component of mature Vo. Voa1’s C-terminal transmembrane α helix is bound inside the proteolipid ring, where it contributes to the stability of the complex. Our structure rationalizes possible mechanisms by which mutations in human Vo can result in disease phenotypes and may thus provide new avenues for therapeutic interventions.},
doi = {10.1016/j.molcel.2018.02.006},
journal = {Molecular Cell},
number = 6,
volume = 69,
place = {United States},
year = {2018},
month = {3}
}

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Works referencing / citing this record:

A cation–π interaction in a transmembrane helix of vacuolar ATPase retains the proton-transporting arginine in a hydrophobic environment
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