Substrate specificity of Staphylococcus aureus cysteine proteases – Staphopains A, B and C

Kalińska, Magdalena; Kantyka, Tomasz; Greenbaum, Doron C.; Larsen, Katrine S.; Władyka, Benedykt; Jabaiah, Abeer; Bogyo, Matthew; Daugherty, Patrick S.; Wysocka, Magdalena; Jaros, M.; Lesner, Adam; Rolka, Krzysztof; Schaschke, Norbert; Stennicke, Henning R.; Dubin, Adam; Potempa, Jan; Dubin, Grzegorz

doi:10.1016/j.biochi.2011.07.020

Cited by 18 publications

(14 citation statements)

References 43 publications

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“…Inhibition of StpC activity during bacterial growth using a specific irreversible inhibitor of cysteine peptidases (E-64) had no effect on the level of BacCH91, suggesting an StpC-independent maturation mechanism. This finding is consistent with the recently reported substrate specificity of StpC for bulky uncharged amino acid residues (Kalinska et al 2012); in turn, the BacCH91 leader peptide is processed after a charged residue (Fig. 3c).…”

Section: Discussionsupporting

confidence: 93%

Isolation, biochemical characterization, and cloning of a bacteriocin from the poultry-associated Staphylococcus aureus strain CH-91

Władyka

Wielebska

Wloka

et al. 2012

Appl Microbiol Biotechnol

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Staphylococcus aureus strain CH-91, isolated from a broiler chicken with atopic dermatitis, has a highly proteolytic phenotype that is correlated with the disease. We describe the isolation and biochemical and molecular characterization of the AI-type lantibiotic BacCH91 from S. aureus CH-91 culture medium. The bacteriocin was purified using a three-stage procedure comprising precipitation with ammonium sulfate, extraction with organic solvents, and reversed-phase HPLC. The BacCH91 peptide is thermostable and highly resistant to cleavage by both prokaryotic and eukaryotic peptidases. The MIC for the Gram-positive bacteria ranged from 2.5 nM for Microococcus luteus through 1.3–6.0 μM for staphylococcal strains up to more than 100 μM for Lactococcus lactis. BacCH91 was ineffective against the Gram-negative strains tested at the maximal concentration (100 μM). The amino acid sequence of BacCH91 is similar to that of epidermin and gallidermin. The encoding gene (bacCH91) occurred in two allelic variants distinguishable in the restriction fragment length polymorphism assay. Variant I, identified in S. aureus CH-91, dominated in S. aureus strains of poultry origin, although strains with variant II were also identified in this group. S. aureus strains of human origin were characterized exclusively by variant II.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-012-4578-y) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 93%

Isolation, biochemical characterization, and cloning of a bacteriocin from the poultry-associated Staphylococcus aureus strain CH-91

Władyka

Wielebska

Wloka

et al. 2012

Appl Microbiol Biotechnol

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“…N‐terminal sequencing revealed that the cleavage product starts with the amino‐acid sequence NH2‐AAP*EPES, indicating that Staphopain A cleaves between Aspartate (D35) and Alanine (A36): LLD↓A (Figure 6B; Supplementary Figure 4). In a recent study, the Staphopain A substrate binding site was mapped using a bacterial cell‐surface display system (CLiPS; Kalińska et al , 2012). This CLiPS study indicated that Staphopain A prefers a Leucine residue for the P2 site of the substrate, corresponding with the Leucine at position 34 in CXCR2.…”

Section: Resultsmentioning

confidence: 99%

“…The CXCR2 1-48 -GB1-His protein was detected using western blotting with an antibody against the His tag. or Ser) position (Kaliń ska et al, 2012). The LLDkA site also contains an Alanine at the P1 0 site.…”

Section: Discussionmentioning

confidence: 99%

Staphylococcus aureusStaphopain A inhibits CXCR2-dependent neutrophil activation and chemotaxis

et al. 2012

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“…This process may be followed by secondary necrosis [85], as we observed LDH release and 7AAD staining. Interestingly, cathepsin B was identified as the closest structural neighbor of staphopains among the eukaryotic enzymes [48] and the overall substrate preferences of ScpA and SspB most closely resembled that of human cathepsin H [86]. Cathepsins are eukaryotic proteases, which are associated with cellular protein turnover, but also induce apoptosis by cleavage of Bid and degradation of the anti-apoptotic Bcl-2 homologues [87–90].…”

Section: Discussionmentioning

confidence: 99%

IntracellularStaphylococcus aureusemploys the cysteine protease staphopain A to induce host cell death in epithelial cells

Stelzner

Hertlein

Sroka

et al. 2020

Preprint

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Staphylococcus aureus is a major human pathogen, which can invade and survive in non-professional and professional phagocytes. Intracellularity is thought to contribute to pathogenicity and persistence of the bacterium. Upon internalization by epithelial cells, cytotoxic S. aureus strains can escape from the phagosome, replicate in the cytosol and induce host cell death. Here, we identified a staphylococcal cysteine protease to induce cell death by intracellular S. aureus after translocation into the host cell cytoplasm. We demonstrated that loss of staphopain A function leads to delayed onset of host cell death and prolonged intracellular replication of S. aureus in epithelial cells. Overexpression of staphopain A in a non-cytotoxic strain facilitated intracellular killing of the host cell even in the absence of detectable intracellular replication. Moreover, staphopain A contributed to efficient colonization of the lung in a mouse pneumonia model. Our study suggests that staphopain A is utilized by S. aureus to mediate escape from the host cell and thus contributes to tissue destruction and dissemination of infection. Author Summary Staphylococcus aureus is a well-known antibiotic-resistant pathogen that emerges in hospital and community settings and can cause a variety of diseases ranging from skin abscesses to lung inflammation and blood poisoning. The bacterium asymptomatically colonizes the upper respiratory tract and skin of about one third of the human population and takes advantage of opportune conditions, like immunodeficiency or breached barriers, to cause infection. Although S. aureus is not regarded as a professional intracellular bacterium, it can be internalized by human cells and subsequently exit the host cells by induction of cell death, which is considered to cause tissue destruction and spread of infection. The bacterial virulence factors and underlying molecular mechanisms involved in the intracellular lifestyle of S. aureus remain largely unknown. We identified a bacterial cysteine protease to contribute to host cell death mediated by intracellular S. aureus. Staphopain A induced killing of the host cell after translocation of the pathogen into the cell cytosol, while bacterial proliferation was not required. Further, the protease enhanced survival of the pathogen during lung infection. These findings reveal a novel, intracellular role for the bacterial protease staphopain A.

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Substrate specificity of Staphylococcus aureus cysteine proteases – Staphopains A, B and C

Cited by 18 publications

References 43 publications

Isolation, biochemical characterization, and cloning of a bacteriocin from the poultry-associated Staphylococcus aureus strain CH-91

Isolation, biochemical characterization, and cloning of a bacteriocin from the poultry-associated Staphylococcus aureus strain CH-91

Staphylococcus aureusStaphopain A inhibits CXCR2-dependent neutrophil activation and chemotaxis

IntracellularStaphylococcus aureusemploys the cysteine protease staphopain A to induce host cell death in epithelial cells

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