Structural insights into recognition of chemokine receptors by Staphylococcus aureus leukotoxins
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Curated by eLife
Evaluation Summary:
This manuscript reports on the interaction of LukE toxin with chemokine receptors. The study is extensive and employs a wide array of tools, although further experimentation would be needed to substantiate the inferences made by the authors. The paper will be of interest to scientists interested in host-pathogen interactions as it delves into understanding the molecular mechanism and interactions of an important toxin interacting with cellular receptors.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)
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Abstract
Staphylococcus aureus (SA) leukocidin ED (LukED) belongs to a family of bicomponent pore forming toxins that play important roles in SA immune evasion and nutrient acquisition. LukED targets specific G protein-coupled chemokine receptors to lyse human erythrocytes (red blood cells) and leukocytes (white blood cells). The first recognition step of receptors is critical for specific cell targeting and lysis. The structural and molecular bases for this mechanism are not well understood but could constitute essential information to guide antibiotic development. Here, we characterized the interaction of LukE with chemokine receptors ACKR1, CCR2, and CCR5 using a combination of structural, pharmacological, and computational approaches. First, crystal structures of LukE in complex with a small molecule mimicking sulfotyrosine side chain (p-cresyl sulfate) and with peptides containing sulfotyrosines issued from receptor sequences revealed the location of receptor sulfotyrosine binding sites in the toxins. Then, by combining previous and novel experimental data with protein docking, classical and accelerated weight histogram (AWH) molecular dynamics we propose models of the ACKR1-LukE and CCR5-LukE complexes. This work provides novel insights into chemokine receptor recognition by leukotoxins and suggests that the conserved sulfotyrosine binding pocket could be a target of choice for future drug development.
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Evaluation Summary:
This manuscript reports on the interaction of LukE toxin with chemokine receptors. The study is extensive and employs a wide array of tools, although further experimentation would be needed to substantiate the inferences made by the authors. The paper will be of interest to scientists interested in host-pathogen interactions as it delves into understanding the molecular mechanism and interactions of an important toxin interacting with cellular receptors.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)
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Reviewer #1 (Public Review):
Lambey et al. used x-ray crystallography, docking, and MD simulations to generate high-quality models of chemokine receptors interacting with Staphylococcus aureus (SA) leukotoxins (LukE). SA leukotoxins cause severe infections and invade host cells by first interacting with chemokine receptors on the host cell. The structural models emerging from this work identify novel interactions sites that are also highly conserved. This work is significant because, in addition to advancing our mechanistic understanding of the recognition of features exploited by LukE, it identified specific sites that we could be target and design inhibitors that prevent leukotoxins from invading host cells.
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Reviewer #2 (Public Review):
The manuscript authored by Lambey et al delves into the interactions of leukotoxins secreted by S.aureus and their interactions with chemokine receptors prior to pore formation. This study uses a wide array of techniques to understand an important step in the host-pathogen interaction that leads to specific targeting of chemokine receptor-containing cells by a staphylococcal leukotoxin component (LukE) which proceeds to cell lysis when the F component for the toxin is available (LukD). The authors use X-ray structures of the LukE subunit (previously determined by Nocadello et al., 2016) in apo state and in complex with the p-cresol sulfate to mimic sulfated tyrosine and N-terminal sulfated tyrosine containing peptides of the CCR2 and ACKR1 receptors to demonstrate the importance of this posttranslational …
Reviewer #2 (Public Review):
The manuscript authored by Lambey et al delves into the interactions of leukotoxins secreted by S.aureus and their interactions with chemokine receptors prior to pore formation. This study uses a wide array of techniques to understand an important step in the host-pathogen interaction that leads to specific targeting of chemokine receptor-containing cells by a staphylococcal leukotoxin component (LukE) which proceeds to cell lysis when the F component for the toxin is available (LukD). The authors use X-ray structures of the LukE subunit (previously determined by Nocadello et al., 2016) in apo state and in complex with the p-cresol sulfate to mimic sulfated tyrosine and N-terminal sulfated tyrosine containing peptides of the CCR2 and ACKR1 receptors to demonstrate the importance of this posttranslational modification (sulfation) in dictating the interactions with the host chemokine receptors. In addition, they perform a very nice FRET experiment to demonstrate the interaction of the LukE to compete the native labeled ligand of the chemokine receptors to demonstrate nanomolar affinity of LukE to ACKR1, CCR2 and CCR5 receptors. They further use docking tools and MD analysis to suggest interaction sites of the LukE toxin subunit with the chemokine receptors.
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Reviewer #3 (Public Review):
Manuscript by Lambey et al attempts to understand structural insights how bacterial toxin LukE binds to cytokine receptors. This is an interesting biological problem to understand, which may facilitate development of new antibiotics. Authors determined two crystal structures of LukE at 1.5 and 1.9 A resolutions and three complexes of the toxin with bound p-cresol sulfate or sulfated peptides derived from cytokine receptors. Based on these data, authors provide novel insights into the recognition of sulfated tyrosine residues in the N-terminal region of cytokine receptors. To understand the overall architecture of LukE-receptor complexes, they performed extensive MD simulations supported by previously reported mutagenesis data. Overall, the models of LukE complexes look reasonable; however, these models lack …
Reviewer #3 (Public Review):
Manuscript by Lambey et al attempts to understand structural insights how bacterial toxin LukE binds to cytokine receptors. This is an interesting biological problem to understand, which may facilitate development of new antibiotics. Authors determined two crystal structures of LukE at 1.5 and 1.9 A resolutions and three complexes of the toxin with bound p-cresol sulfate or sulfated peptides derived from cytokine receptors. Based on these data, authors provide novel insights into the recognition of sulfated tyrosine residues in the N-terminal region of cytokine receptors. To understand the overall architecture of LukE-receptor complexes, they performed extensive MD simulations supported by previously reported mutagenesis data. Overall, the models of LukE complexes look reasonable; however, these models lack sufficient experimental validation. In addition, more data are needed to map the binding site of N-terminal fragment of cytokine receptors in solution. Binding sites of p-cresol sulfate and peptides with sulfated tyrosines have been obtained from crystal soaking experiments and may be biased by crystal packing. As presented, data are preliminary and need to be strengthened by additional biophysical/structural studies.
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