Lipopolysaccharide (LPS) transport to the outer membrane (OM) is a crucial step in the biogenesis of microbial surface defenses. Although many features of the translocation mechanism have been elucidated, molecular details of LPS insertion via the LPS transport (Lpt) OM protein LptDE remain elusive. Here, we integrate native MS with hydrogen–deuterium exchange MS and molecular dynamics simulations to investigate the influence of substrate and peptide binding on the conformational dynamics of LptDE. Our data reveal that LPS induces opening of the LptD β-taco domain, coupled with conformational changes on β-strands adjacent to the putative lateral exit gate. Conversely, an antimicrobial peptide, thanatin, stabilizes the β-taco, thereby preventing LPS transport. Our results illustrate that LPS insertion into the OM relies on concerted opening movements of both the β-barrel and β-taco domains of LptD, and suggest a means for developing antimicrobial therapeutics targeting this essential process in Gram-negative ESKAPE pathogens.

Dynamics of an LPS translocon induced by substrate and an antimicrobial peptide / Fiorentino, F.; Sauer, J. B.; Qiu, X.; Corey, R. A.; Cassidy, C. K.; Mynors-Wallis, B.; Mehmood, S.; Bolla, J. R.; Stansfeld, P. J.; Robinson, C. V.. - In: NATURE CHEMICAL BIOLOGY. - ISSN 1552-4450. - 17:2(2021), pp. 187-195. [10.1038/s41589-020-00694-2]

Dynamics of an LPS translocon induced by substrate and an antimicrobial peptide

Fiorentino F.
Primo
;
2021

Abstract

Lipopolysaccharide (LPS) transport to the outer membrane (OM) is a crucial step in the biogenesis of microbial surface defenses. Although many features of the translocation mechanism have been elucidated, molecular details of LPS insertion via the LPS transport (Lpt) OM protein LptDE remain elusive. Here, we integrate native MS with hydrogen–deuterium exchange MS and molecular dynamics simulations to investigate the influence of substrate and peptide binding on the conformational dynamics of LptDE. Our data reveal that LPS induces opening of the LptD β-taco domain, coupled with conformational changes on β-strands adjacent to the putative lateral exit gate. Conversely, an antimicrobial peptide, thanatin, stabilizes the β-taco, thereby preventing LPS transport. Our results illustrate that LPS insertion into the OM relies on concerted opening movements of both the β-barrel and β-taco domains of LptD, and suggest a means for developing antimicrobial therapeutics targeting this essential process in Gram-negative ESKAPE pathogens.
2021
Anti-Bacterial Agents; Antimicrobial Cationic Peptides; Bacterial Outer Membrane Proteins; Carbohydrate Conformation; Drug Resistance, Bacterial; Gram-Negative Bacteria; Klebsiella pneumoniae; Lipopolysaccharides; Mass Spectrometry; Models, Molecular; Molecular Dynamics Simulation; Translocation, Genetic
01 Pubblicazione su rivista::01a Articolo in rivista
Dynamics of an LPS translocon induced by substrate and an antimicrobial peptide / Fiorentino, F.; Sauer, J. B.; Qiu, X.; Corey, R. A.; Cassidy, C. K.; Mynors-Wallis, B.; Mehmood, S.; Bolla, J. R.; Stansfeld, P. J.; Robinson, C. V.. - In: NATURE CHEMICAL BIOLOGY. - ISSN 1552-4450. - 17:2(2021), pp. 187-195. [10.1038/s41589-020-00694-2]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1585455
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