Protease IV, a Unique Extracellular Protease and Virulence Factor from Pseudomonas aeruginosa

Engel, Lee S.; Hill, James M.; Caballero, Armando R.; Green, Linda C.; O’Callaghan, Richard J.

doi:10.1074/jbc.273.27.16792

Cited by 152 publications

(169 citation statements)

References 27 publications

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“…followed by uptake of the bacteria in these organs. Clearance through this mechanism should decrease the rate at which otherwise free (not E-bound) bacteria can invade other organs and tissues, including the lungs, which are particularly susceptible to PAO1 (54,55). The decreased pathology associated with the lungs in the HP-treated monkeys (see above) is consistent with this hypothesis.…”

Section: Effects Of Hp On Bacterial Clearancesupporting

confidence: 77%

Targeting of Pseudomonas aeruginosa in the Bloodstream with Bispecific Monoclonal Antibodies

Lindorfer

Nardin

Foley³

et al. 2001

The Journal of Immunology

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We examined the ability of a bispecific mAb reagent, consisting of a mAb specific for the primate erythrocyte complement receptor cross-linked with an anti-bacterial mAb, to target bacteria in the bloodstream in an acute infusion model in monkeys. In vitro studies demonstrated a variable level of complement-mediated binding (immune adherence) of Pseudomonas aeruginosa (strain PAO1) to primate E in serum. In vivo experiments in animals depleted of complement revealed that binding of bacteria to E was <1% before administration of the bispecific reagent, but within 5 min of its infusion, >99% of the bacteria bound to E. In complement-replete monkeys, a variable fraction of infused bacteria bound to E. This finding may have significant implications in the interpretation of animal models and in the understanding of bacteremias in humans. Treatment of these complement-replete monkeys with the bispecific reagent led to >99% binding of bacteria to E. Twenty-four-hour survival studies were conducted; several clinical parameters, including the degree of lung damage, cytokine levels, and liver enzymes in the circulation, indicate that the bispecific mAb reagent provides a degree of protection against the bacterial challenge.

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Section: Effects Of Hp On Bacterial Clearancesupporting

confidence: 77%

Targeting of Pseudomonas aeruginosa in the Bloodstream with Bispecific Monoclonal Antibodies

Lindorfer

Nardin

Foley³

et al. 2001

The Journal of Immunology

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“…Gelatin zymography techniques based on those of Twining et al (1993) were employed to determine protease IV activity in the isolates. Cell-free culture supernatants were preincubated with or without the protease IV specific inhibitor N-a-rtosyl-L-lysine chloromethyl ketone (TLCK, Sigma-Aldrich) (Engel et al, 1998). TLCK was prepared as a stock solution of 10 mM in 5 mM HCl.…”

Section: Methodsmentioning

confidence: 99%

“…Mature protease IV degrades important host immunological proteins such as complement and IgG (Engel et al, 1998). Protease IV also compromises the integrity of structural proteins such as elastin (Wilderman et al, 2001), causing tissue damage and facilitating bacterial infection.…”

Section: Introductionmentioning

confidence: 99%

Characterization of protease IV expression in Pseudomonas aeruginosa clinical isolates

Conibear

Willcox

Flanagan

et al. 2012

Journal of Medical Microbiology

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Expression of protease IV by Pseudomonas aeruginosa during ocular infections contributes significantly to tissue damage. However, several P. aeruginosa strains isolated from ocular infections or inflammatory events produce very low levels of protease IV. The aim of the present study was to characterize, genetically and phenotypically, the presence and expression of the protease IV gene in a group of clinical isolates that cause adverse ocular events of varying degrees, and to elucidate the possible control mechanisms of expression associated with this virulence factor. Protease IV gene sequences from seven clinical isolates of P. aeruginosa were determined and compared to P. aeruginosa strains PAO1 and PA103-29. Production and enzyme activity of protease IV were measured in test strains and compared to that of quorumsensing gene (lasRI) mutants and the expression of other virulence factors. Protease IV gene sequence similarities between the isolates were 97.5-99.5 %. The strains were classified into two distinct phylogenetic groups that correlated with the presence of exo-enzymes from type three secretion systems (TTSS). Protease IV concentrations produced by PAODlasRI mutants and the two clinical isolates with a lasRI gene deficiency were restored to levels comparable to strain PAO1 following complementation of the quorum-sensing gene deficiencies. The protease IV gene is highly conserved in P. aeruginosa clinical isolates that cause a range of adverse ocular events. Observed variations within the gene sequence appear to correlate with presence of specific TTSS genes. Protease IV expression was shown to be regulated by the Las quorum-sensing system. INTRODUCTIONInflammation and infection in the cornea (keratitis) arising from Pseudomonas aeruginosa infection can lead rapidly to scarring and loss of sight unless prompt and targeted treatment is initiated (Sankaridurg et al., 2000). P. aeruginosa virulence is attributed, in part, to the production of several destructive proteins and stimulation of host immune response in the ocular tissues (Hazlett, 2002). The virulence factors most commonly associated with P. aeruginosa-induced ocular damage are exo-enzymes S (exoS) and U (exoU) (Fleiszig et al., 1997), elastase (lasB) (Kessler & Blumberg, 1987), alkaline protease (aprA) (Twining et al., 1993) and protease IV (piv) (O'Callaghan et al., 1996). Another protease, P. aeruginosa small protease (PASP), can also cause rapid corneal damage (Marquart et al., 2005). Other important pathogenic mechanisms include the genes involved in translocation and activation of virulence proteins via the type II (xcp) and type III secretory pathways (Sandkvist, 2001;Yahr et al., 1997).Protease IV is a lysine-specific endoprotease and a significant virulence factor for pathogenesis of P. aeruginosa in the eye and lung (Caballero et al., 2004; Engel et al., 1997;Wilderman et al., 2001). Mature protease IV degrades important host immunological proteins such as complement and IgG (Engel et al., 1998). Protease IV also compromises the inte...

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“…LasA protease can also nick elastin at certain Gly-Gly sequences (9,10), an activity that increases the susceptibility of elastin to other proteases and contributes to the elastinolytic potential of P. aeruginosa (9,11,12). The fourth endopeptidase secreted by P. aeruginosa (lysine-specific endopeptidase; protease IV) cleaves peptide bonds on the carboxyl side of lysine residues in peptides and proteins and can act on a number of host proteins including complement components, IgG, and fibrinogen (13)(14)(15). LasD, a secreted protein first described as a second staphylolytic protease of P. aeruginosa (16), is apparently not a protease but may function as a chitin-binding protein (17).…”

mentioning

confidence: 99%

A Secreted Aminopeptidase of Pseudomonas aeruginosa

Cahan¹,

Axelrad²,

Safrin³

et al. 2001

Journal of Biological Chemistry

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Using leucine-p-nitroanilide (Leu-pNA) as a substrate, we demonstrated aminopeptidase activity in the culture filtrates of several Pseudomonas aeruginosa strains. The aminopeptidase was partially purified by DEAE-cellulose chromatography and found to be heat stable. The apparent molecular mass of the enzyme was ϳ56 kDa; hence, it was designated AP 56 . Heating (70°C) of the partially purified aminopeptidase preparations led to the conversion of AP 56 to a ϳ28-kDa protein (AP 28 ) that retained enzyme activity, a reaction that depended on elastase (LasB). The pH optimum for Leu-pNA hydrolysis by AP 28 was 8.5. This activity was inhibited by Zn chelators but not by inhibitors of serine-or thiol-proteases, suggesting that AP 28 is a Zn-dependent enzyme. Of several amino acid p-nitroanilide derivatives examined, Leu-pNA was the preferred substrate. The sequences of the first 20 residues of AP 56 and AP 28 were determined. A search of the P. aeruginosa genomic data base revealed a perfect match of these sequences with positions 39 -58 and 273-291, respectively, in a 536-amino acid residue open reading frame predicted to encode an aminopeptidase. A search for sequence similarities with other proteins revealed 52% identity with Streptomyces griseus aminopeptidase, ϳ35% identity with Saccharomyces cerevisiae aminopeptidase Y and a hypothetical aminopeptidase from Bacillus subtilis, and 29 -32% with Aeromonas caviae, Vibrio proteolyticus, and Vibrio cholerae aminopeptidases. The residues potentially involved in zinc coordination were conserved in all these proteins. Thus, P. aeruginosa aminopeptidase may belong to the same family (M28) of metalloproteases.

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Protease IV, a Unique Extracellular Protease and Virulence Factor from Pseudomonas aeruginosa

Cited by 152 publications

References 27 publications

Targeting of Pseudomonas aeruginosa in the Bloodstream with Bispecific Monoclonal Antibodies

Targeting of Pseudomonas aeruginosa in the Bloodstream with Bispecific Monoclonal Antibodies

Characterization of protease IV expression in Pseudomonas aeruginosa clinical isolates

A Secreted Aminopeptidase of Pseudomonas aeruginosa

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