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

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

doi:10.1101/2023.01.02.522452

bioRxiv · 10.1101/2023.01.02.522452

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

Yang, Y.·Chen, H.·Corey, R.·Morales, V.·Quentin, Y.·Froment, C.·Caumont-Sarcos, A.·Albenne, C.·Burlet-Schiltz, O.·Ranava, D.·Stansfeld, P. J.·Marcoux, J.·Ieva, R.

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Abstract

Insertion of lipopolysaccharide (LPS) into the outer membrane (OM) of Gram-negative bacteria is mediated by a druggable OM translocon consisting of a {beta}-barrel membrane protein, LptD, and a lipoprotein, LptE. The {beta}-barrel assembly machinery (BAM) assembles LptD together with LptE to form a plug-and-barrel structure. In the enterobacterium Escherichia coli, formation of two native disulfide bonds in LptD controls LPS translocon activation. Here we report the discovery of LptM (formerly YifL), a conserved lipoprotein that assembles together with LptD and LptE at the BAM complex. We demonstrate that LptM stabilizes a conformation of LptD that can efficiently acquire native disulfide bonds and be released as mature LPS translocon by the BAM complex. Inactivation of LptM causes the accumulation of non-natively oxidized LptD, making 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 LptM facilitates oxidative maturation of LptD by mimicking LPS binding, thereby activating the LPS translocon.

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Yang, Y., Chen, H., Corey, R., Morales, V., Quentin, Y., Froment, C., Caumont-Sarcos, A., Albenne, C., Burlet-Schiltz, O., Ranava, D., Stansfeld, P. J., Marcoux, J., Ieva, R.. 2023-01-03. LptM promotes oxidative maturation of the lipopolysaccharide translocon by substrate binding mimicry.. https://doi.org/10.1101/2023.01.02.522452

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