Bovine F1Fo ATP synthase monomers bend the lipid bilayer in 2D membrane crystals

  1. Chimari Jiko
  2. Karen M Davies
  3. Kyoko Shinzawa-Itoh
  4. Kazutoshi Tani
  5. Shintaro Maeda
  6. Deryck J Mills
  7. Tomitake Tsukihara
  8. Yoshinori Fujiyoshi
  9. Werner Kühlbrandt
  10. Christoph Gerle  Is a corresponding author
  1. Osaka University, Japan
  2. Max Planck Institute of Biophysics, Germany
  3. University of Hyogo, Japan
  4. Nagoya University, Japan

Abstract

We have used a combination of electron cryo-tomography, subtomogram averaging and electron crystallographic image processing to analyze the structure of intact bovine F1Fo ATP synthase in 2D membrane crystals. ATPase assays and mass spectrometry analysis of the 2D crystals confirmed the enzyme complex was complete and active. The structure of the matrix-exposed region was determined at 24 Å resolution by subtomogram averaging, and repositioned into the tomographic volume to reveal the crystal packing. F1Fo ATP synthase complexes are inclined by 16{degree sign} relative to the crystal plane, resulting in a zigzag topology of the membrane and indicating that monomeric bovine heart F1Fo ATP synthase by itself is sufficient to deform lipid bilayers. This local membrane curvature is likely to be instrumental in the formation of ATP synthase dimers and dimer rows, and thus for the shaping of mitochondrial cristae.

Article and author information

Author details

  1. Chimari Jiko

    Institute for Protein Research, Osaka University, Osaka, Japan
    Competing interests
    No competing interests declared.
  2. Karen M Davies

    Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt am Main, Germany
    Competing interests
    No competing interests declared.
  3. Kyoko Shinzawa-Itoh

    Picobiology Institute, Department of Life Science, Graduate School of Life Science, University of Hyogo, Kamigori, Japan
    Competing interests
    No competing interests declared.
  4. Kazutoshi Tani

    Cellular and Structural Physiology Institute, Nagoya University, Nagoya, Japan
    Competing interests
    No competing interests declared.
  5. Shintaro Maeda

    Picobiology Institute, Department of Life Science, Graduate School of Life Science, University of Hyogo, Kamigori, Japan
    Competing interests
    No competing interests declared.
  6. Deryck J Mills

    Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt am Main, Germany
    Competing interests
    No competing interests declared.
  7. Tomitake Tsukihara

    Picobiology Institute, Department of Life Science, Graduate School of Life Science, University of Hyogo, Kamigori, Japan
    Competing interests
    No competing interests declared.
  8. Yoshinori Fujiyoshi

    Cellular and Structural Physiology Institute, Nagoya University, Nagoya, Japan
    Competing interests
    No competing interests declared.
  9. Werner Kühlbrandt

    Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt am Main, Germany
    Competing interests
    Werner Kühlbrandt, Reviewing editor, eLife.
  10. Christoph Gerle

    Picobiology Institute, Department of Life Science, Graduate School of Life Science, University of Hyogo, Kamigori, Japan
    For correspondence
    gerle.christoph@gmail.com
    Competing interests
    No competing interests declared.

Reviewing Editor

  1. Sjors HW Scheres, Medical Research Council Laboratory of Molecular Biology, United Kingdom

Version history

  1. Received: December 16, 2014
  2. Accepted: March 26, 2015
  3. Accepted Manuscript published: March 27, 2015 (version 1)
  4. Version of Record published: April 29, 2015 (version 2)

Copyright

© 2015, Jiko et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,218
    views
  • 732
    downloads
  • 67
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Chimari Jiko
  2. Karen M Davies
  3. Kyoko Shinzawa-Itoh
  4. Kazutoshi Tani
  5. Shintaro Maeda
  6. Deryck J Mills
  7. Tomitake Tsukihara
  8. Yoshinori Fujiyoshi
  9. Werner Kühlbrandt
  10. Christoph Gerle
(2015)
Bovine F1Fo ATP synthase monomers bend the lipid bilayer in 2D membrane crystals
eLife 4:e06119.
https://doi.org/10.7554/eLife.06119

Share this article

https://doi.org/10.7554/eLife.06119

Further reading

    1. Structural Biology and Molecular Biophysics
    Marco van den Noort, Panagiotis Drougkas ... Bert Poolman
    Research Article

    Bacteria utilize various strategies to prevent internal dehydration during hypertonic stress. A common approach to countering the effects of the stress is to import compatible solutes such as glycine betaine, leading to simultaneous passive water fluxes following the osmotic gradient. OpuA from Lactococcus lactis is a type I ABC-importer that uses two substrate-binding domains (SBDs) to capture extracellular glycine betaine and deliver the substrate to the transmembrane domains for subsequent transport. OpuA senses osmotic stress via changes in the internal ionic strength and is furthermore regulated by the 2nd messenger cyclic-di-AMP. We now show, by means of solution-based single-molecule FRET and analysis with multi-parameter photon-by-photon hidden Markov modeling, that the SBDs transiently interact in an ionic strength-dependent manner. The smFRET data are in accordance with the apparent cooperativity in transport and supported by new cryo-EM data of OpuA. We propose that the physical interactions between SBDs and cooperativity in substrate delivery are part of the transport mechanism.

    1. Structural Biology and Molecular Biophysics
    Xiao-Ru Chen, Karuna Dixit ... Tatyana I Igumenova
    Research Article

    Regulated hydrolysis of the phosphoinositide phosphatidylinositol(4,5)-bis-phosphate to diacylglycerol and inositol-1,4,5-P3 defines a major eukaryotic pathway for translation of extracellular cues to intracellular signaling circuits. Members of the lipid-activated protein kinase C isoenzyme family (PKCs) play central roles in this signaling circuit. One of the regulatory mechanisms employed to downregulate stimulated PKC activity is via a proteasome-dependent degradation pathway that is potentiated by peptidyl-prolyl isomerase Pin1. Here, we show that contrary to prevailing models, Pin1 does not regulate conventional PKC isoforms α and βII via a canonical cis-trans isomerization of the peptidyl-prolyl bond. Rather, Pin1 acts as a PKC binding partner that controls PKC activity via sequestration of the C-terminal tail of the kinase. The high-resolution structure of full-length Pin1 complexed to the C-terminal tail of PKCβII reveals that a novel bivalent interaction mode underlies the non-catalytic mode of Pin1 action. Specifically, Pin1 adopts a conformation in which it uses the WW and PPIase domains to engage two conserved phosphorylated PKC motifs, the turn motif and hydrophobic motif, respectively. Hydrophobic motif is a non-canonical Pin1-interacting element. The structural information combined with the results of extensive binding studies and experiments in cultured cells suggest that non-catalytic mechanisms represent unappreciated modes of Pin1-mediated regulation of AGC kinases and other key enzymes/substrates.