LptM promotes oxidative maturation of the lipopolysaccharide translocon by substrate binding mimicry

Y Yang; H Chen; RA Corey; V Morales; Y Quentin; C Froment; A Caumont-Sarcos; C Albenne; O Burlet-Schiltz; D Ranava; PJ Stansfeld; J Marcoux; R Ieva

doi:10.1038/s41467-023-42007-w

LptM promotes oxidative maturation of the lipopolysaccharide translocon by substrate binding mimicry

Y Yang, H Chen, RA Corey, V Morales, Y Quentin, C Froment, A Caumont-Sarcos, C Albenne, O Burlet-Schiltz, D Ranava, PJ Stansfeld, J Marcoux, R Ieva^*

^*Corresponding author for this work

School of Physiology, Pharmacology & Neuroscience

Research output: Contribution to journal › Article (Academic Journal) › peer-review

6 Citations (Scopus)

Abstract

Insertion of lipopolysaccharide (LPS) into the bacterial outer membrane (OM) is mediated by a druggable OM translocon consisting of a β-barrel membrane protein, LptD, and a lipoprotein, LptE. The β-barrel assembly machinery (BAM) assembles LptD together with LptE at the OM. In the enterobacterium Escherichia coli, formation of two native disulfide bonds in LptD controls translocon activation. Here we report the discovery of LptM (formerly YifL), a lipoprotein conserved in Enterobacteriaceae, that assembles together with LptD and LptE at the BAM complex. LptM stabilizes a conformation of LptD that can efficiently acquire native disulfide bonds, whereas its inactivation makes disulfide bond isomerization by DsbC become essential for viability. Our structural prediction and biochemical analyses indicate that LptM binds to sites in both LptD and LptE that are proposed to coordinate LPS insertion into the OM. These results suggest that, by mimicking LPS binding, LptM facilitates oxidative maturation of LptD, thereby activating the LPS translocon.

Original language	English
Article number	6368
Number of pages	15
Journal	Nature Communications
Volume	14
DOIs	https://doi.org/10.1038/s41467-023-42007-w
Publication status	Published - 11 Oct 2023

Bibliographical note

Funding Information:
Research in R.I.’s lab was funded by the CNRS (ATIP-Avenir grant to R.I.), the China Scholarship Council fellowships to Y.Y. and H.C. and the FRM postdoctoral fellowship to D.R. HDX-MS experiments were supported by the French Ministry of Research (Investissements d’Avenir Program, Proteomics French Infrastructure, ANR-10-INBS-08 to O. B.-S.) and the Région Midi Pyrénées to O.B.-S. Research in P.J.S.’s lab was funded by Wellcome (208361/Z/17/Z) and BBSRC (BB/P01948X/1, BB/R002517/1 and BB/S003339/1). This project made use of time on ARCHER2 and JADE2 granted via the UK High-End Computing Consortium for Biomolecular Simulation, HECBioSim (http://hecbiosim.ac.uk), supported by EPSRC (grant no. EP/R029407/1). This project also used Athena and Sulis at HPC Midlands + , which were funded by the EPSRC on grants EP/P020232/1 and EP/T022108/1. We thank the LMGM, Toulouse, for access to the E. coli Keio collection. We thank the IPBS, Toulouse, for access to the MALDI-TOF/TOF mass spectrometer. We thank the University of Warwick Scientific Computing Research Technology Platform for computational access.

Funding Information:
Research in R.I.’s lab was funded by the CNRS (ATIP-Avenir grant to R.I.), the China Scholarship Council fellowships to Y.Y. and H.C. and the FRM postdoctoral fellowship to D.R. HDX-MS experiments were supported by the French Ministry of Research (Investissements d’Avenir Program, Proteomics French Infrastructure, ANR-10-INBS-08 to O. B.-S.) and the Région Midi Pyrénées to O.B.-S. Research in P.J.S.’s lab was funded by Wellcome (208361/Z/17/Z) and BBSRC (BB/P01948X/1, BB/R002517/1 and BB/S003339/1). This project made use of time on ARCHER2 and JADE2 granted via the UK High-End Computing Consortium for Biomolecular Simulation, HECBioSim ( http://hecbiosim.ac.uk ), supported by EPSRC (grant no. EP/R029407/1). This project also used Athena and Sulis at HPC Midlands + , which were funded by the EPSRC on grants EP/P020232/1 and EP/T022108/1. We thank the LMGM, Toulouse, for access to the E. coli Keio collection. We thank the IPBS, Toulouse, for access to the MALDI-TOF/TOF mass spectrometer. We thank the University of Warwick Scientific Computing Research Technology Platform for computational access.

Publisher Copyright:
© 2023, Springer Nature Limited.

Keywords

membrane protein
Lipid

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10.1038/s41467-023-42007-w

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title = "LptM promotes oxidative maturation of the lipopolysaccharide translocon by substrate binding mimicry",

abstract = "Insertion of lipopolysaccharide (LPS) into the bacterial outer membrane (OM) is mediated by a druggable OM translocon consisting of a β-barrel membrane protein, LptD, and a lipoprotein, LptE. The β-barrel assembly machinery (BAM) assembles LptD together with LptE at the OM. In the enterobacterium Escherichia coli, formation of two native disulfide bonds in LptD controls translocon activation. Here we report the discovery of LptM (formerly YifL), a lipoprotein conserved in Enterobacteriaceae, that assembles together with LptD and LptE at the BAM complex. LptM stabilizes a conformation of LptD that can efficiently acquire native disulfide bonds, whereas its inactivation makes disulfide bond isomerization by DsbC become essential for viability. Our structural prediction and biochemical analyses indicate that LptM binds to sites in both LptD and LptE that are proposed to coordinate LPS insertion into the OM. These results suggest that, by mimicking LPS binding, LptM facilitates oxidative maturation of LptD, thereby activating the LPS translocon.",

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author = "Y Yang and H Chen and RA Corey and V Morales and Y Quentin and C Froment and A Caumont-Sarcos and C Albenne and O Burlet-Schiltz and D Ranava and PJ Stansfeld and J Marcoux and R Ieva",

note = "Funding Information: Research in R.I.{\textquoteright}s lab was funded by the CNRS (ATIP-Avenir grant to R.I.), the China Scholarship Council fellowships to Y.Y. and H.C. and the FRM postdoctoral fellowship to D.R. HDX-MS experiments were supported by the French Ministry of Research (Investissements d{\textquoteright}Avenir Program, Proteomics French Infrastructure, ANR-10-INBS-08 to O. B.-S.) and the R{\'e}gion Midi Pyr{\'e}n{\'e}es to O.B.-S. Research in P.J.S.{\textquoteright}s lab was funded by Wellcome (208361/Z/17/Z) and BBSRC (BB/P01948X/1, BB/R002517/1 and BB/S003339/1). This project made use of time on ARCHER2 and JADE2 granted via the UK High-End Computing Consortium for Biomolecular Simulation, HECBioSim (http://hecbiosim.ac.uk), supported by EPSRC (grant no. EP/R029407/1). This project also used Athena and Sulis at HPC Midlands + , which were funded by the EPSRC on grants EP/P020232/1 and EP/T022108/1. We thank the LMGM, Toulouse, for access to the E. coli Keio collection. We thank the IPBS, Toulouse, for access to the MALDI-TOF/TOF mass spectrometer. We thank the University of Warwick Scientific Computing Research Technology Platform for computational access. Funding Information: Research in R.I.{\textquoteright}s lab was funded by the CNRS (ATIP-Avenir grant to R.I.), the China Scholarship Council fellowships to Y.Y. and H.C. and the FRM postdoctoral fellowship to D.R. HDX-MS experiments were supported by the French Ministry of Research (Investissements d{\textquoteright}Avenir Program, Proteomics French Infrastructure, ANR-10-INBS-08 to O. B.-S.) and the R{\'e}gion Midi Pyr{\'e}n{\'e}es to O.B.-S. Research in P.J.S.{\textquoteright}s lab was funded by Wellcome (208361/Z/17/Z) and BBSRC (BB/P01948X/1, BB/R002517/1 and BB/S003339/1). This project made use of time on ARCHER2 and JADE2 granted via the UK High-End Computing Consortium for Biomolecular Simulation, HECBioSim ( http://hecbiosim.ac.uk ), supported by EPSRC (grant no. EP/R029407/1). This project also used Athena and Sulis at HPC Midlands + , which were funded by the EPSRC on grants EP/P020232/1 and EP/T022108/1. We thank the LMGM, Toulouse, for access to the E. coli Keio collection. We thank the IPBS, Toulouse, for access to the MALDI-TOF/TOF mass spectrometer. We thank the University of Warwick Scientific Computing Research Technology Platform for computational access. Publisher Copyright: {\textcopyright} 2023, Springer Nature Limited.",

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doi = "10.1038/s41467-023-42007-w",

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T1 - LptM promotes oxidative maturation of the lipopolysaccharide translocon by substrate binding mimicry

AU - Yang, Y

AU - Chen, H

AU - Corey, RA

AU - Morales, V

AU - Quentin, Y

AU - Froment, C

AU - Caumont-Sarcos, A

AU - Albenne, C

AU - Burlet-Schiltz, O

AU - Ranava, D

AU - Stansfeld, PJ

AU - Marcoux, J

AU - Ieva, R

N1 - Funding Information: Research in R.I.’s lab was funded by the CNRS (ATIP-Avenir grant to R.I.), the China Scholarship Council fellowships to Y.Y. and H.C. and the FRM postdoctoral fellowship to D.R. HDX-MS experiments were supported by the French Ministry of Research (Investissements d’Avenir Program, Proteomics French Infrastructure, ANR-10-INBS-08 to O. B.-S.) and the Région Midi Pyrénées to O.B.-S. Research in P.J.S.’s lab was funded by Wellcome (208361/Z/17/Z) and BBSRC (BB/P01948X/1, BB/R002517/1 and BB/S003339/1). This project made use of time on ARCHER2 and JADE2 granted via the UK High-End Computing Consortium for Biomolecular Simulation, HECBioSim (http://hecbiosim.ac.uk), supported by EPSRC (grant no. EP/R029407/1). This project also used Athena and Sulis at HPC Midlands + , which were funded by the EPSRC on grants EP/P020232/1 and EP/T022108/1. We thank the LMGM, Toulouse, for access to the E. coli Keio collection. We thank the IPBS, Toulouse, for access to the MALDI-TOF/TOF mass spectrometer. We thank the University of Warwick Scientific Computing Research Technology Platform for computational access. Funding Information: Research in R.I.’s lab was funded by the CNRS (ATIP-Avenir grant to R.I.), the China Scholarship Council fellowships to Y.Y. and H.C. and the FRM postdoctoral fellowship to D.R. HDX-MS experiments were supported by the French Ministry of Research (Investissements d’Avenir Program, Proteomics French Infrastructure, ANR-10-INBS-08 to O. B.-S.) and the Région Midi Pyrénées to O.B.-S. Research in P.J.S.’s lab was funded by Wellcome (208361/Z/17/Z) and BBSRC (BB/P01948X/1, BB/R002517/1 and BB/S003339/1). This project made use of time on ARCHER2 and JADE2 granted via the UK High-End Computing Consortium for Biomolecular Simulation, HECBioSim ( http://hecbiosim.ac.uk ), supported by EPSRC (grant no. EP/R029407/1). This project also used Athena and Sulis at HPC Midlands + , which were funded by the EPSRC on grants EP/P020232/1 and EP/T022108/1. We thank the LMGM, Toulouse, for access to the E. coli Keio collection. We thank the IPBS, Toulouse, for access to the MALDI-TOF/TOF mass spectrometer. We thank the University of Warwick Scientific Computing Research Technology Platform for computational access. Publisher Copyright: © 2023, Springer Nature Limited.

PY - 2023/10/11

Y1 - 2023/10/11

N2 - Insertion of lipopolysaccharide (LPS) into the bacterial outer membrane (OM) is mediated by a druggable OM translocon consisting of a β-barrel membrane protein, LptD, and a lipoprotein, LptE. The β-barrel assembly machinery (BAM) assembles LptD together with LptE at the OM. In the enterobacterium Escherichia coli, formation of two native disulfide bonds in LptD controls translocon activation. Here we report the discovery of LptM (formerly YifL), a lipoprotein conserved in Enterobacteriaceae, that assembles together with LptD and LptE at the BAM complex. LptM stabilizes a conformation of LptD that can efficiently acquire native disulfide bonds, whereas its inactivation makes disulfide bond isomerization by DsbC become essential for viability. Our structural prediction and biochemical analyses indicate that LptM binds to sites in both LptD and LptE that are proposed to coordinate LPS insertion into the OM. These results suggest that, by mimicking LPS binding, LptM facilitates oxidative maturation of LptD, thereby activating the LPS translocon.

AB - Insertion of lipopolysaccharide (LPS) into the bacterial outer membrane (OM) is mediated by a druggable OM translocon consisting of a β-barrel membrane protein, LptD, and a lipoprotein, LptE. The β-barrel assembly machinery (BAM) assembles LptD together with LptE at the OM. In the enterobacterium Escherichia coli, formation of two native disulfide bonds in LptD controls translocon activation. Here we report the discovery of LptM (formerly YifL), a lipoprotein conserved in Enterobacteriaceae, that assembles together with LptD and LptE at the BAM complex. LptM stabilizes a conformation of LptD that can efficiently acquire native disulfide bonds, whereas its inactivation makes disulfide bond isomerization by DsbC become essential for viability. Our structural prediction and biochemical analyses indicate that LptM binds to sites in both LptD and LptE that are proposed to coordinate LPS insertion into the OM. These results suggest that, by mimicking LPS binding, LptM facilitates oxidative maturation of LptD, thereby activating the LPS translocon.

KW - membrane protein

KW - Lipid

UR - http://europepmc.org/abstract/med/37821449

U2 - 10.1038/s41467-023-42007-w

DO - 10.1038/s41467-023-42007-w

M3 - Article (Academic Journal)

C2 - 37821449

SN - 2041-1723

VL - 14

JO - Nature Communications

JF - Nature Communications

M1 - 6368

ER -

LptM promotes oxidative maturation of the lipopolysaccharide translocon by substrate binding mimicry

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