Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH

Abstract Wall teichoic acid (WTA) is a polyol phosphate polymer that covalently decorates peptidoglycan of gram-positive bacteria, including Staphylococcus aureus. Central to WTA biosynthesis is flipping of lipid-linked precursors across the cell membrane by TarGH, a type V ABC transporter. Here, we...

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Main Authors: Franco K. K. Li, Shaun C. Peters, Liam J. Worrall, Tianjun Sun, Jinhong Hu, Marija Vuckovic, Maya Farha, Armando Palacios, Nathanael A. Caveney, Eric D. Brown, Natalie C. J. Strynadka
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58202-w
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author Franco K. K. Li
Shaun C. Peters
Liam J. Worrall
Tianjun Sun
Jinhong Hu
Marija Vuckovic
Maya Farha
Armando Palacios
Nathanael A. Caveney
Eric D. Brown
Natalie C. J. Strynadka
author_facet Franco K. K. Li
Shaun C. Peters
Liam J. Worrall
Tianjun Sun
Jinhong Hu
Marija Vuckovic
Maya Farha
Armando Palacios
Nathanael A. Caveney
Eric D. Brown
Natalie C. J. Strynadka
author_sort Franco K. K. Li
collection DOAJ
description Abstract Wall teichoic acid (WTA) is a polyol phosphate polymer that covalently decorates peptidoglycan of gram-positive bacteria, including Staphylococcus aureus. Central to WTA biosynthesis is flipping of lipid-linked precursors across the cell membrane by TarGH, a type V ABC transporter. Here, we present cryo-EM structures of S. aureus TarGH in the presence of targocil-II, a promising small-molecule lead with β-lactam antibiotic synergistic action. Targocil-II binds to the extracellular dimerisation interface of TarG, we suggest mimicking flipped but not yet released substrate. In absence of targocil-II and in complex with ATP analogue ATPγS, determined at 2.3 Å resolution, the ATPase active site is allosterically inhibited. This is due to a so far undescribed D-loop conformation, potentially minimizing spurious ATP hydrolysis in the absence of substrate. Targocil-II binding comparatively causes local and remote conformational changes through to the TarH active site, with the D-loop now optimal for ATP hydrolysis. These structures suggest an ability to modulate ATP hydrolysis in a WTA substrate dependent manner and a jammed ATPase cycle as the basis of the observed inhibition by targocil-II. The molecular insights provide an unprecedented basis for development of TarGH targeted therapeutics for treatment of multidrug-resistant S. aureus and other gram-positive bacterial infections.
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spelling doaj-art-fe910a7763b7406da2d583ad88a1bda22025-08-20T01:54:30ZengNature PortfolioNature Communications2041-17232025-04-0116111410.1038/s41467-025-58202-wCryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGHFranco K. K. Li0Shaun C. Peters1Liam J. Worrall2Tianjun Sun3Jinhong Hu4Marija Vuckovic5Maya Farha6Armando Palacios7Nathanael A. Caveney8Eric D. Brown9Natalie C. J. Strynadka10Department of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Biomedical Sciences, McMaster UniversityDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaDepartment of Biochemistry and Biomedical Sciences, McMaster UniversityDepartment of Biochemistry and Molecular Biology and the Centre for Blood Research, University of British ColumbiaAbstract Wall teichoic acid (WTA) is a polyol phosphate polymer that covalently decorates peptidoglycan of gram-positive bacteria, including Staphylococcus aureus. Central to WTA biosynthesis is flipping of lipid-linked precursors across the cell membrane by TarGH, a type V ABC transporter. Here, we present cryo-EM structures of S. aureus TarGH in the presence of targocil-II, a promising small-molecule lead with β-lactam antibiotic synergistic action. Targocil-II binds to the extracellular dimerisation interface of TarG, we suggest mimicking flipped but not yet released substrate. In absence of targocil-II and in complex with ATP analogue ATPγS, determined at 2.3 Å resolution, the ATPase active site is allosterically inhibited. This is due to a so far undescribed D-loop conformation, potentially minimizing spurious ATP hydrolysis in the absence of substrate. Targocil-II binding comparatively causes local and remote conformational changes through to the TarH active site, with the D-loop now optimal for ATP hydrolysis. These structures suggest an ability to modulate ATP hydrolysis in a WTA substrate dependent manner and a jammed ATPase cycle as the basis of the observed inhibition by targocil-II. The molecular insights provide an unprecedented basis for development of TarGH targeted therapeutics for treatment of multidrug-resistant S. aureus and other gram-positive bacterial infections.https://doi.org/10.1038/s41467-025-58202-w
spellingShingle Franco K. K. Li
Shaun C. Peters
Liam J. Worrall
Tianjun Sun
Jinhong Hu
Marija Vuckovic
Maya Farha
Armando Palacios
Nathanael A. Caveney
Eric D. Brown
Natalie C. J. Strynadka
Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH
Nature Communications
title Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH
title_full Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH
title_fullStr Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH
title_full_unstemmed Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH
title_short Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH
title_sort cryo em analyses unveil details of mechanism and targocil ii mediated inhibition of s aureus wta transporter targh
url https://doi.org/10.1038/s41467-025-58202-w
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