Structural basis of interprotein electron transfer in bacterial sulfite oxidation

  1. Aaron P McGrath
  2. Elise L Laming
  3. G Patricia Casas Garcia
  4. Marc Kvansakul
  5. J Mitchell Guss
  6. Jill Trewhella
  7. Benoit Calmes
  8. Paul V Bernhardt
  9. Graeme R Hanson
  10. Ulrike Kappler
  11. Megan J Maher  Is a corresponding author
  1. University of California, San Diego, United States
  2. The Victor Chang Cardiac Research Institute, Australia
  3. La Trobe University, Australia
  4. University of Sydney, Australia
  5. University of Queensland, Australia

Abstract

Interprotein electron transfer underpins the essential processes of life and relies on the formation of specific, yet transient protein-protein interactions. In biological systems, the detoxification of sulfite is catalyzed by the sulfite-oxidizing enzymes (SOEs), which interact with an electron acceptor for catalytic turnover. Here, we report the structural and functional analyses of the SOE SorT from Sinorhizobium meliloti and its cognate electron acceptor SorU. Kinetic and thermodynamic analyses of the SorT/SorU interaction showed the complex is dynamic in solution, and that the proteins interact with Kd = 13.5 {plus minus} 0.8 βM. The crystal structures of the oxidized SorT and SorU both in isolation and in complex, reveal the interface to be remarkably electrostatic, with an unusually large number of direct hydrogen bonding interactions. The assembly of the complex is accompanied by an adjustment in the structure of SorU and conformational sampling provides a mechanism for dissociation of the SorT/SorU assembly.

Article and author information

Author details

  1. Aaron P McGrath

    Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Elise L Laming

    The Victor Chang Cardiac Research Institute, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  3. G Patricia Casas Garcia

    La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia
    Competing interests
    The authors declare that no competing interests exist.
  4. Marc Kvansakul

    La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia
    Competing interests
    The authors declare that no competing interests exist.
  5. J Mitchell Guss

    School of Molecular Bioscience, University of Sydney, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  6. Jill Trewhella

    School of Molecular Bioscience, University of Sydney, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  7. Benoit Calmes

    Centre for Metals in Biology, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  8. Paul V Bernhardt

    Centre for Metals in Biology, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  9. Graeme R Hanson

    Centre for Metals in Biology, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  10. Ulrike Kappler

    Centre for Metals in Biology, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  11. Megan J Maher

    La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia
    For correspondence
    m.maher@latrobe.edu.au
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Michael A Marletta, University of California, Berkeley, United States

Version history

  1. Received: May 28, 2015
  2. Accepted: November 12, 2015
  3. Accepted Manuscript published: December 19, 2015 (version 1)
  4. Accepted Manuscript updated: December 23, 2015 (version 2)
  5. Version of Record published: February 4, 2016 (version 3)

Copyright

© 2015, McGrath 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

  • 1,326
    views
  • 287
    downloads
  • 18
    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. Aaron P McGrath
  2. Elise L Laming
  3. G Patricia Casas Garcia
  4. Marc Kvansakul
  5. J Mitchell Guss
  6. Jill Trewhella
  7. Benoit Calmes
  8. Paul V Bernhardt
  9. Graeme R Hanson
  10. Ulrike Kappler
  11. Megan J Maher
(2015)
Structural basis of interprotein electron transfer in bacterial sulfite oxidation
eLife 4:e09066.
https://doi.org/10.7554/eLife.09066

Share this article

https://doi.org/10.7554/eLife.09066

Further reading

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    Amy H Andreotti, Volker Dötsch
    Editorial

    The articles in this special issue highlight how modern cellular, biochemical, biophysical and computational techniques are allowing deeper and more detailed studies of allosteric kinase regulation.

    1. Developmental Biology
    2. Structural Biology and Molecular Biophysics
    Samuel C Griffiths, Jia Tan ... Hsin-Yi Henry Ho
    Research Article Updated

    The receptor tyrosine kinase ROR2 mediates noncanonical WNT5A signaling to orchestrate tissue morphogenetic processes, and dysfunction of the pathway causes Robinow syndrome, brachydactyly B, and metastatic diseases. The domain(s) and mechanisms required for ROR2 function, however, remain unclear. We solved the crystal structure of the extracellular cysteine-rich (CRD) and Kringle (Kr) domains of ROR2 and found that, unlike other CRDs, the ROR2 CRD lacks the signature hydrophobic pocket that binds lipids/lipid-modified proteins, such as WNTs, suggesting a novel mechanism of ligand reception. Functionally, we showed that the ROR2 CRD, but not other domains, is required and minimally sufficient to promote WNT5A signaling, and Robinow mutations in the CRD and the adjacent Kr impair ROR2 secretion and function. Moreover, using function-activating and -perturbing antibodies against the Frizzled (FZ) family of WNT receptors, we demonstrate the involvement of FZ in WNT5A-ROR signaling. Thus, ROR2 acts via its CRD to potentiate the function of a receptor super-complex that includes FZ to transduce WNT5A signals.