The hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switch

Abstract The peptidoglycan (PG) cell wall is the primary protective layer of bacteria, making the process of PG synthesis a key antibiotic target. Class A penicillin-binding proteins (aPBPs) are a family of conserved and ubiquitous PG synthases that fortify and repair the PG matrix. In gram-negative...

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Main Authors: Irina Shlosman, Andrea Vettiger, Thomas G. Bernhardt, Andrew C. Kruse, Joseph J. Loparo
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62051-y
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author Irina Shlosman
Andrea Vettiger
Thomas G. Bernhardt
Andrew C. Kruse
Joseph J. Loparo
author_facet Irina Shlosman
Andrea Vettiger
Thomas G. Bernhardt
Andrew C. Kruse
Joseph J. Loparo
author_sort Irina Shlosman
collection DOAJ
description Abstract The peptidoglycan (PG) cell wall is the primary protective layer of bacteria, making the process of PG synthesis a key antibiotic target. Class A penicillin-binding proteins (aPBPs) are a family of conserved and ubiquitous PG synthases that fortify and repair the PG matrix. In gram-negative bacteria, these enzymes are regulated by outer-membrane tethered lipoproteins. However, the molecular mechanism by which lipoproteins coordinate the spatial recruitment and enzymatic activation of aPBPs remains unclear. Here we use single-molecule FRET and single-particle tracking in E. coli to show that a prototypical lipoprotein activator LpoB triggers site-specific PG synthesis by PBP1b through conformational rearrangements. Once synthesis is initiated, LpoB affinity for PBP1b dramatically decreases and it dissociates from the synthesizing enzyme. Our results suggest that transient allosteric coupling between PBP1b and LpoB directs PG synthesis to areas of low peptidoglycan density, while simultaneously facilitating efficient lipoprotein redistribution to other sites in need of fortification.
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institution Kabale University
issn 2041-1723
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spelling doaj-art-b7d3e60183604eb7a216ec82f70420092025-08-20T04:03:03ZengNature PortfolioNature Communications2041-17232025-07-0116111510.1038/s41467-025-62051-yThe hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switchIrina Shlosman0Andrea Vettiger1Thomas G. Bernhardt2Andrew C. Kruse3Joseph J. Loparo4Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical SchoolDepartment of Fundamental Microbiology, Université de LausanneDepartment of Microbiology, Blavatnik Institute, Harvard Medical SchoolDepartment of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical SchoolDepartment of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical SchoolAbstract The peptidoglycan (PG) cell wall is the primary protective layer of bacteria, making the process of PG synthesis a key antibiotic target. Class A penicillin-binding proteins (aPBPs) are a family of conserved and ubiquitous PG synthases that fortify and repair the PG matrix. In gram-negative bacteria, these enzymes are regulated by outer-membrane tethered lipoproteins. However, the molecular mechanism by which lipoproteins coordinate the spatial recruitment and enzymatic activation of aPBPs remains unclear. Here we use single-molecule FRET and single-particle tracking in E. coli to show that a prototypical lipoprotein activator LpoB triggers site-specific PG synthesis by PBP1b through conformational rearrangements. Once synthesis is initiated, LpoB affinity for PBP1b dramatically decreases and it dissociates from the synthesizing enzyme. Our results suggest that transient allosteric coupling between PBP1b and LpoB directs PG synthesis to areas of low peptidoglycan density, while simultaneously facilitating efficient lipoprotein redistribution to other sites in need of fortification.https://doi.org/10.1038/s41467-025-62051-y
spellingShingle Irina Shlosman
Andrea Vettiger
Thomas G. Bernhardt
Andrew C. Kruse
Joseph J. Loparo
The hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switch
Nature Communications
title The hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switch
title_full The hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switch
title_fullStr The hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switch
title_full_unstemmed The hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switch
title_short The hit-and-run of cell wall synthesis: LpoB transiently binds and activates PBP1b through a conserved allosteric switch
title_sort hit and run of cell wall synthesis lpob transiently binds and activates pbp1b through a conserved allosteric switch
url https://doi.org/10.1038/s41467-025-62051-y
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