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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]
+ Show Author Affiliations
  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.
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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.
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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.
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@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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DOI:  10.1016/j.molcel.2018.02.006
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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
journal, September 2018

  • Hohlweg, Walter; Wagner, Gabriel E.; Hofbauer, Harald F.
  • Journal of Biological Chemistry, Vol. 293, Issue 49
  • DOI: 10.1074/jbc.ra118.005276

A cation–π interaction in a transmembrane helix of vacuolar ATPase retains the proton-transporting arginine in a hydrophobic environment
journal, September 2018

  • Hohlweg, Walter; Wagner, Gabriel E.; Hofbauer, Harald F.
  • Journal of Biological Chemistry, Vol. 293, Issue 49
  • DOI: 10.1074/jbc.ra118.005276

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  • Renaud, Jean-Paul; Chari, Ashwin; Ciferri, Claudio
  • Nature Reviews Drug Discovery, Vol. 17, Issue 7
  • DOI: 10.1038/nrd.2018.77

Unusual features of the c-ring of F1FO ATP synthases
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Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae
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  • Vasanthakumar, Thamiya; Bueler, Stephanie A.; Wu, Di
  • Proceedings of the National Academy of Sciences, Vol. 116, Issue 15
  • DOI: 10.1073/pnas.1814818116

Atp6ap2 deletion causes extensive vacuolation that consumes the insulin content of pancreatic β cells
journal, September 2019

  • Binger, Katrina J.; Neukam, Martin; Tattikota, Sudhir Gopal
  • Proceedings of the National Academy of Sciences, Vol. 116, Issue 40
  • DOI: 10.1073/pnas.1903678116

Functional reconstitution of vacuolar H + -ATPase from V o proton channel and mutant V 1 -ATPase provides insight into the mechanism of reversible disassembly
journal, February 2019

  • Sharma, Stuti; Oot, Rebecca A.; Khan, Md Murad
  • Journal of Biological Chemistry, Vol. 294, Issue 16
  • DOI: 10.1074/jbc.ra119.007577

Interaction of the late endo-lysosomal lipid PI(3,5)P2 with the Vph1 isoform of yeast V-ATPase increases its activity and cellular stress tolerance
journal, April 2019

  • Banerjee, Subhrajit; Clapp, Kaitlyn; Tarsio, Maureen
  • Journal of Biological Chemistry, Vol. 294, Issue 23
  • DOI: 10.1074/jbc.ra119.008552

ATP6AP2 functions as a V-ATPase assembly factor in the endoplasmic reticulum
journal, September 2018

  • Guida, Maria Clara; Hermle, Tobias; Graham, Laurie A.
  • Molecular Biology of the Cell, Vol. 29, Issue 18
  • DOI: 10.1091/mbc.e18-04-0234

Cryo-EM structure of the mammalian ATP synthase tetramer bound with inhibitory protein IF1
journal, June 2019

Structure and conformational plasticity of the intact Thermus thermophilus V/A-type ATPase
journal, August 2019

Regulation of V-ATPase Assembly in Nutrient Sensing and Function of V-ATPases in Breast Cancer Metastasis
journal, July 2018

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