Substitution of cysteine 192 in a highly conserved Streptococcus pyogenes extracellular cysteine protease (interleukin 1beta convertase) alters proteolytic activity and ablates zymogen processing

Musser, James M.; Stockbauer, Kathryn E.; Kapur, Vivek; Rudgers, Gary W.

doi:10.1128/iai.64.6.1913-1917.1996

Cited by 51 publications

(25 citation statements)

References 47 publications

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“…Site-specific mutagenesis confirmed the critical role of the Cys at position 192 (C192), the His at position 340 (H340), and the Trp at position 357 (W357), for enzymatic activity. Replacing C192 with Ser or H340 with Ala produced a proteolytically inactive mutant protein while replacement of W357 with Ala resulted in a 50 000-fold decrease in enzyme activity, confirming that these residues are important for SpeBm activity (Elliott, 1945;Liu et al, 1963;Liu, 1967;Musser et al, 1996;Gubba et al, 2000;Chen et al, 2003).…”

Section: Structure and Processing Of Spebzmentioning

confidence: 85%

From transcription to activation: how group A streptococcus, the flesh‐eating pathogen, regulates SpeB cysteine protease production

Carroll

Musser²

2011

Molecular Microbiology

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SummaryStreptococcal pyrogenic exotoxin B (SpeB) is a protease secreted by group A streptococci and known to degrade a wide range of host and GAS proteins in vitro. Although the role of SpeB in GAS infection is debated, recent evidence has conclusively demonstrated that SpeB is critical for the pathogenesis of severe invasive disease caused by GAS. Genetic inactivation of the speB gene results in significantly decreased virulence in a necrotizing fasciitis model of infection. Production of fully active SpeB by GAS is extremely complex. Following transcription and translation the SpeB protein is secreted as an inactive zymogen, which is autocatalytically processed through a series of intermediates to form an active protease. Each step from transcription to protease activation is tightly controlled and regulated by the bacterial cell reflecting the critical role played by this virulence factor in GAS infection. Here we review the molecular aspects of SpeB production by GAS from transcription to activation and the multiple layers of control involved.

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Section: Structure and Processing Of Spebzmentioning

confidence: 85%

From transcription to activation: how group A streptococcus, the flesh‐eating pathogen, regulates SpeB cysteine protease production

Carroll

Musser²

2011

Molecular Microbiology

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“…Under reducing conditions, the proprotein can undergo autocatalytic cleavage to the mature active 28 kDa form. Consistent with a requirement for reducing conditions, mutation of the single cysteine residue of the proprotein blocks the ability of the resulting mutant protein to undergo autocatalytic processing (Musser et al ., 1996). This level of processing suggests the involvement of a complex secretion mechanism.…”

Section: Introductionmentioning

confidence: 97%

A role for Trigger Factor and an Rgg-like regulator in the transcription, secretion and processing of the cysteine proteinase ofStreptococcus pyogenes

1998

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The ability of numerous microorganisms to cause disease relies upon the highly regulated expression of secreted proteinases. In this study, mutagenesis with a novel derivative of Tn4001 was used to identify genes required for the expression of the secreted cysteine proteinase (SCP) of the pathogenic Gram-positive bacterium Streptococcus pyogenes. Designated as Rop loci (regulation of proteinase), ropB is a rgg-like transcriptional activator required for transcription of the gene which encodes the proteinase. In contrast, ropA contributes post-transcriptionally to the secretion and processing of SCP and encodes a homologue of Trigger Factor, a peptidyl-prolyl isomerase and putative chaparone which is highly conserved in most bacterial species, but of unknown function. Analysis of additional ropA mutants demonstrated that RopA acts both to assist in targeting SCP to the secretory pathway and to promote the ability of the proprotein to establish an active conformation upon secretion. This latter function was dependent upon the peptidyl-prolyl isomerase domain of RopA and mutants that lacked this domain exhibited a bipartite deficiency manifested as a kinetic defect in autologous processing of the proprotein to the mature proteinase, and as a catalytic defect in the mature proteinase. These results provide insight into the function of Trigger Factor, the regulation of proteinase activity and the mechanism of secretion in Gram-positive bacteria.

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“…Among the secreted factors, the cysteine protease SpeB has been suggested as contributing to the invasiveness of GAS (Talkington et al, 1993;Raeder and Boyle, 1995;Shanley et al, 1996). Although initially characterized as a mitogenic factor (Hauser and Schlievert, 1990), the molecule has recently been shown to be a cysteine protease (Chaussee et al, 1993;Ohara-Nemoto et al, 1994;Musser et al, 1996). It was found to cleave human serum proteins like pre-IL-1b (Kapur et al, 1993a), fibronectin (Kapur et al, 1993b), human cell-surface proteins like the urokinase-receptor (Wolf et al, 1994), and streptococcal surface proteins, M protein and C5a peptidase (Berge and Björck, 1995).…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of group A streptococcal (GAS) oligopeptide permease (Opp) and its effect on cysteine protease production

Podbielski¹,

Pohl²,

Woischnik³

et al. 1996

Molecular Microbiology

115

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Bacterial oligopeptide permeases are membrane-associated complexes of five proteins belonging to the ABC-transporter family, which have been found to be involved in obtaining nutrients, cell-wall metabolism, competence, and adherence to host cells. A lambda library of the strain CS101 group A streptococcal (GAS) genome was used to sequence 10,192 bp containing the five genes oppA to oppF of the GAS opp operon. The deduced amino acid sequences exhibited 50-84% homology to pneumococcal AmiA to AmiF sequences. The operon organization of the five genes was confirmed by transcriptional analysis and an additional shorter oppA transcript was detected. Insertional inactivation was used to create serotype M49 strains which did not express either the oppA gene or the ATPase genes, oppD and oppF. The mutation in oppA confirmed that the additional shorter oppA transcript originated from the opp operon and was probably due to an intra-operon transcription terminator site located downstream of oppA. While growth kinetics, binding of serum proteins, and attachment to eukaryotic cells were unaffected, the oppD/F mutants showed reduced production of the cysteine protease, SpeB, and a change in the pattern of secreted proteins. Thus, the GAS opp operon appears to contribute to both protease production and export/processing of secreted proteins.

show abstract

Substitution of cysteine 192 in a highly conserved Streptococcus pyogenes extracellular cysteine protease (interleukin 1beta convertase) alters proteolytic activity and ablates zymogen processing

Cited by 51 publications

References 47 publications

From transcription to activation: how group A streptococcus, the flesh‐eating pathogen, regulates SpeB cysteine protease production

From transcription to activation: how group A streptococcus, the flesh‐eating pathogen, regulates SpeB cysteine protease production

A role for Trigger Factor and an Rgg-like regulator in the transcription, secretion and processing of the cysteine proteinase ofStreptococcus pyogenes

Molecular characterization of group A streptococcal (GAS) oligopeptide permease (Opp) and its effect on cysteine protease production

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