Functional and Structural Characterization of Spl Proteases from Staphylococcus aureus

Popowicz, Grzegorz M.; Dubin, Grzegorz; Steć-Niemczyk, Justyna; Czarny, Anna; Dubin, Adam; Potempa, Jan; Holak, T.A.

doi:10.1016/j.jmb.2006.01.098

Cited by 51 publications

(74 citation statements)

References 36 publications

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“…It was proposed that contact with substrate containing peptides would trigger a conformational change of the flexible loop, allowing the catalytic histidine to rotate into the active site. Studies with recombinant Spl proteins also supported the contention that even a single additional amino acid at the N terminus would be sufficient to maintain zymogen status (46).…”

Section: Discussionmentioning

confidence: 83%

“…The exfoliative toxins are glutamyl endopeptidases that primarily cleave a single host protein, desmoglein (49), whereas the Spl proteins are encoded by tandem gene repeats in a genomic island structure and are either very restricted in their substrate specificity or could not be demonstrated to have protease activity (46,50). Presumably because they are restricted in their substrate specificity, there is no need for an intramolecular inhibitor.…”

Section: Discussionmentioning

confidence: 99%

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Activation of the SspA Serine Protease Zymogen of Staphylococcus aureus Proceeds through Unique Variations of a Trypsinogen-like Mechanism and Is Dependent on Both Autocatalytic and Metalloprotease-specific Processing

Nickerson

Prasad

Jacob

et al. 2007

Journal of Biological Chemistry

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The serine and cysteine proteases SspA and SspB of Staphylococcus aureus are secreted as inactive zymogens, zSspA and zSspB. Mature SspA is a trypsin-like glutamyl endopeptidase and is required to activate zSspB. Although a metalloprotease Aureolysin (Aur) is in turn thought to contribute to activation of zSspA, a specific role has not been demonstrated. We found that pre-zSspA is processed by signal peptidase at ANA 29 Binding of glutamate within the active site of zSspA is energetically unfavorable, but glutamine fits into the primary specificity pocket and is predicted to hydrogen bond to Thr 232 proximal to Ser 237 , permitting autocatalytic cleavage of the glutamine-rich propeptide segment. These and other observations suggest that zSspA is activated through a trypsinogen-like mechanism where supplementary features of the propeptide must be sequentially processed in the correct order to allow efficient activation.Staphylococcus aureus can colonize and infect virtually every tissue and organ system of the body (1, 2). A key factor in these defining traits is its ability to sustain bacteremia and adhere to tissues. Consequently it has adapted to growth in blood by subversion of plasma proteins (3) and the tissue extracellular matrix (4). Members of the MSCRAMM (microbial extracellular matrix-binding protein) family of adhesion proteins bind extracellular matrix ligands such as collagen, fibronectin, and fibrinogen (4) of which the latter two are also abundant in plasma. Manipulation of other plasma proteins such as IgG and von Willebrand factor is facilitated by staphylococcal Protein A (5), whereas coagulase promotes fibrin clot formation by activation of prothrombin (6), and staphylokinase activates plasminogen (7) to facilitate fibrin clot dissolution. Our studies on secreted proteases are also supportive of S. aureus as a paradigm for the manipulation of plasma and coagulation proteins.The SspA 4 glutamyl endopeptidase, also known as V8 protease, moderates adhesion of S. aureus to fibronectin by degrading cell surface fibronectin-binding proteins in which there is a high glutamic acid content, including a conserved motif that is essential for ligand binding (8, 9). SspA is expressed in an operon with a cysteine protease, SspB, and activates the zSspB zymogen by removing its N-terminal propeptide (10). Mature 20-kDa SspB mimics plasma serine proteases as noted by its ability to (i) convert high molecular weight kininogen into heavy and light chains, (ii) hydrolyze the Bz-Pro-Phe-Arg substrate of kallikrein, a serine protease that processes single chain kininogen by excision of the vasoactive peptide bradykinin (RPPGFSPFR), (iii) process the N terminus of the fibrinogen ␤-chain at the same site as plasmin, and (iv) remove the N-terminal domain of fibronectin with a specificity equivalent to plasminogen activator (10). We have also found that antibodies to SspB are produced when mice are challenged with hypervirulent strains of community-acquired methicillin-resistant S. aureus and that SspA and SspB are r...

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Section: Discussionmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Activation of the SspA Serine Protease Zymogen of Staphylococcus aureus Proceeds through Unique Variations of a Trypsinogen-like Mechanism and Is Dependent on Both Autocatalytic and Metalloprotease-specific Processing

Nickerson

Prasad

Jacob

et al. 2007

Journal of Biological Chemistry

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“…Construction of Expression Vectors-A gene encoding signal peptide containing SplB (sp-SplB) and its fragment encoding mature SplB protease were PCR-amplified from Staphylococcus aureus 8325-4 genomic DNA and cloned into pGEX-5T (11) using XhoI and BamHI restriction sites, resulting in pSplB(T) and p(sp-SplB(T)) partially as described previously (12). The thrombin site was exchanged in both plasmids into that recognized by factor Xa by site-directed mutagenesis, resulting in pSplB(X) and p(sp-SplB(X)).…”

Section: Methodsmentioning

confidence: 99%

“…Of the recombinant protease variants tested, only those with the N terminus corresponding to the natural protein but not those containing amino acids artificially added in cloning, demonstrated activity in a zymography assay (12). Using a more sensitive and quantitative assay, we show here that the activity of a full-length, 36-amino acid signal peptide containing SplB protease variant (sp-SplB) is not fully abolished, but decreased by more than an order of magnitude compared with that of the mature protease (SplB).…”

Section: Precise Processing At the N Terminus Regulates The Activity Ofmentioning

confidence: 99%

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Staphylococcal SplB Serine Protease Utilizes a Novel Molecular Mechanism of Activation

Pustelny

Zdzalik

Stach

et al. 2014

Journal of Biological Chemistry

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Background: Activation of SplB protease requires precise N-terminal processing, but the molecular mechanism remains unknown. Results:The new N-terminal Glu-1 forms a distinctive H-bond network essential for full catalytic activity. Conclusion: Changes in protein dynamics rather than a direct effect on the active site are of crucial importance in SplB activation. Significance: A novel serine protease activation mechanism was uncovered.

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Promising abilities of mercapto‐degrading Staphylococcus capitis strain SH6 in both crude oil and waste motor oil as sole carbon and energy sources: its biosurfactant production and preliminary characterization

Chebbi

Hentati

Cheffi

et al. 2018

J of Chemical Tech & Biotech

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BACKGROUND It was shown previously that a Staphylococcus capitis strain SH6 was able to degrade several malodorous mercaptans and simultaneously reduce the surface tension. RESULTS This present work revealed the capacity of strain SH6 to grow on various hydrocarbons, used as the only carbon and energy sources. Based on GC–MS analyses, the substantial ability to degrade up to 45% and 64% of aliphatic hydrocarbons (n‐alkanes) of crude oil and waste motor oil, respectively, after 30 days of incubation at 37 °C and 180 rpm, was shown. The properties of biosurfactant produced by strain SH6 grown on different oil substrates (diesel oil and waste motor oil) were studied. Biosurfactants exhibited enhanced emulsification capacities and significant stabilities over a wide range of salinity (20–150 g L‐1), temperature (–20–100 °C), and pH , and also the ability to remove crude oil from contaminated soils. Their critical micelle concentrations (CMC) were of 800 mg L‐1. FTIR analyses suggested the lipopetide nature of biosurfactants. CONCLUSION The stabilities of biosurfactants over a wide pH range, high temperatures and variable concentrations of salt, as well as emulsifying properties, suggest potential applications in bioremediation processes. © 2017 Society of Chemical Industry

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Functional and Structural Characterization of Spl Proteases from Staphylococcus aureus

Cited by 51 publications

References 36 publications

Activation of the SspA Serine Protease Zymogen of Staphylococcus aureus Proceeds through Unique Variations of a Trypsinogen-like Mechanism and Is Dependent on Both Autocatalytic and Metalloprotease-specific Processing

Activation of the SspA Serine Protease Zymogen of Staphylococcus aureus Proceeds through Unique Variations of a Trypsinogen-like Mechanism and Is Dependent on Both Autocatalytic and Metalloprotease-specific Processing

Staphylococcal SplB Serine Protease Utilizes a Novel Molecular Mechanism of Activation

Promising abilities of mercapto‐degrading Staphylococcus capitis strain SH6 in both crude oil and waste motor oil as sole carbon and energy sources: its biosurfactant production and preliminary characterization

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