The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteome

Surface-located and secreted virulence factors of the Gram-positive bacterial pathogen Staphylococcus aureus are key drivers for infection of the human host. Proteolytic enzymes may contribute to virulence by breaking primary barriers and host immune defenses. Therefore, the objective of our study w...

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Main Authors: Xiaofang Li, Sandra Maaß, Borja Ferrero-Bordera, Zhichao Zhang, Min Wang, Eric Sietsema, Lei Liu, Madeleine Divinagracia, Jan Maarten van Dijl, Girbe Buist
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Virulence
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Online Access:https://www.tandfonline.com/doi/10.1080/21505594.2025.2514790
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author Xiaofang Li
Sandra Maaß
Borja Ferrero-Bordera
Zhichao Zhang
Min Wang
Eric Sietsema
Lei Liu
Madeleine Divinagracia
Jan Maarten van Dijl
Girbe Buist
author_facet Xiaofang Li
Sandra Maaß
Borja Ferrero-Bordera
Zhichao Zhang
Min Wang
Eric Sietsema
Lei Liu
Madeleine Divinagracia
Jan Maarten van Dijl
Girbe Buist
author_sort Xiaofang Li
collection DOAJ
description Surface-located and secreted virulence factors of the Gram-positive bacterial pathogen Staphylococcus aureus are key drivers for infection of the human host. Proteolytic enzymes may contribute to virulence by breaking primary barriers and host immune defenses. Therefore, the objective of our study was to chart the contributions of different proteases to S. aureus virulence, and to assess their roles in shaping the staphylococcal surface proteome (the “surfacome”) and extracellular proteome (the “secretome”). To this end, we applied 12 protease mutants of the S. aureus USA300 lineage. Four mutants lacking the metalloprotease aureolysin (Aur), the cysteine proteases staphopain A (ScpA) and SspB, or the serine protease SspA displayed enhanced cytotoxicity toward human lung epithelial cells, showing that they serve to suppress virulence. Profiling of the surfacomes and secretomes of the four mutants allowed correlation of their increased cytotoxicity to altered virulence factor profiles. Furthermore, enhanced levels of virulence factors were detected in the mutants’ surfacomes, which was shown to be relevant as all four mutants displayed enhanced lung epithelial cell invasion. Enhanced levels of cytoplasmic and membrane proteins in the mutant’s surfacomes showed that Aur, ScpA, SspA and SspB set limits to autolysis by reducing the levels of peptidoglycan hydrolases. We conclude that Aur, ScpA, SspA and SspB have key roles in shaping the surfacome and secretome of S. aureus, thereby controlling the virulence of this major pathogen. This implies that novel antimicrobial agents or vaccines should not target these proteases.
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spelling doaj-art-0cb08f7815bd4e4d9cd895e059b9aecd2025-08-20T02:23:11ZengTaylor & Francis GroupVirulence2150-55942150-56082025-12-0116110.1080/21505594.2025.2514790The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteomeXiaofang Li0Sandra Maaß1Borja Ferrero-Bordera2Zhichao Zhang3Min Wang4Eric Sietsema5Lei Liu6Madeleine Divinagracia7Jan Maarten van Dijl8Girbe Buist9Department of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsDepartment of Microbial Proteomics, University of Greifswald, Centre of Functional Genomics of Microbes, Institute of Microbiology, Greifswald, GermanyDepartment of Microbial Proteomics, University of Greifswald, Centre of Functional Genomics of Microbes, Institute of Microbiology, Greifswald, GermanyDepartment of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsDepartment of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsDepartment of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsDepartment of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsDepartment of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsDepartment of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsDepartment of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, the NetherlandsSurface-located and secreted virulence factors of the Gram-positive bacterial pathogen Staphylococcus aureus are key drivers for infection of the human host. Proteolytic enzymes may contribute to virulence by breaking primary barriers and host immune defenses. Therefore, the objective of our study was to chart the contributions of different proteases to S. aureus virulence, and to assess their roles in shaping the staphylococcal surface proteome (the “surfacome”) and extracellular proteome (the “secretome”). To this end, we applied 12 protease mutants of the S. aureus USA300 lineage. Four mutants lacking the metalloprotease aureolysin (Aur), the cysteine proteases staphopain A (ScpA) and SspB, or the serine protease SspA displayed enhanced cytotoxicity toward human lung epithelial cells, showing that they serve to suppress virulence. Profiling of the surfacomes and secretomes of the four mutants allowed correlation of their increased cytotoxicity to altered virulence factor profiles. Furthermore, enhanced levels of virulence factors were detected in the mutants’ surfacomes, which was shown to be relevant as all four mutants displayed enhanced lung epithelial cell invasion. Enhanced levels of cytoplasmic and membrane proteins in the mutant’s surfacomes showed that Aur, ScpA, SspA and SspB set limits to autolysis by reducing the levels of peptidoglycan hydrolases. We conclude that Aur, ScpA, SspA and SspB have key roles in shaping the surfacome and secretome of S. aureus, thereby controlling the virulence of this major pathogen. This implies that novel antimicrobial agents or vaccines should not target these proteases.https://www.tandfonline.com/doi/10.1080/21505594.2025.2514790Staphylococcus aureusvirulenceproteasespeptidoglycan hydrolasecell lysis
spellingShingle Xiaofang Li
Sandra Maaß
Borja Ferrero-Bordera
Zhichao Zhang
Min Wang
Eric Sietsema
Lei Liu
Madeleine Divinagracia
Jan Maarten van Dijl
Girbe Buist
The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteome
Virulence
Staphylococcus aureus
virulence
proteases
peptidoglycan hydrolase
cell lysis
title The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteome
title_full The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteome
title_fullStr The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteome
title_full_unstemmed The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteome
title_short The secreted proteases aur, scpA, sspA and sspB suppress the virulence of Staphylococcus aureus USA300 by shaping the extracellular proteome
title_sort secreted proteases aur scpa sspa and sspb suppress the virulence of staphylococcus aureus usa300 by shaping the extracellular proteome
topic Staphylococcus aureus
virulence
proteases
peptidoglycan hydrolase
cell lysis
url https://www.tandfonline.com/doi/10.1080/21505594.2025.2514790
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