Activation of human pro-urokinase by unrelated proteases secreted by <i>Pseudomonas aeruginosa</i>

Beaufort, Nathalie; Seweryn, Paulina; Bentzmann, Sophie de; Tang, Aihua; Kellermann, Josef; Grebenchtchikov, Nicolaï; Schmitt, Manfred; Sommerhoff, Christian P.; Pidard, Dominique; Magdolen, Viktor

doi:10.1042/bj20091806

Cited by 18 publications

(18 citation statements)

References 51 publications

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“…Degradation of PAI-1 has earlier been observed with the serine protease subtilisin NAT of Bacillus subtilis and the thermolysin-like metalloproteinases aureolysin of Staphylococcus aureus and LasB of Pseudomonas aeruginosa (2,3,7,65). These are extracellularly secreted enzymes, and their cleavage patterns with PAI-1 and plasminogen are different from those of Pla and PgtE.…”

Section: Discussionmentioning

confidence: 99%

“…These are extracellularly secreted enzymes, and their cleavage patterns with PAI-1 and plasminogen are different from those of Pla and PgtE. S. aureus and P. aeruginosa are severe pathogens which show multiple interactions with the human fibrinolytic system (2,6,34,39,61), and LasB is an acknowledged virulence factor in lung and skin infections. A functional difference with pathogenetic potential is that the aureolysin/LasB metalloproteinases cleave plasminogen at the amino-terminal region and at S441-V442, which results in miniplasminogen that is not enzymatically active but can be activated by tPA (2, 3), whereas Pla/PgtE cleave plasminogen at R561-V562, which generates active plasmin.…”

Section: Discussionmentioning

confidence: 99%

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The Omptins ofYersinia pestisandSalmonella entericaCleave the Reactive Center Loop of Plasminogen Activator Inhibitor 1

et al. 2010

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Plasminogen activator inhibitor 1 (PAI-1) is a serine protease inhibitor (serpin) and a key molecule that regulates fibrinolysis by inactivating human plasminogen activators. Here we show that two important human pathogens, the plague bacterium Yersinia pestis and the enteropathogen Salmonella enterica serovar Typhimurium, inactivate PAI-1 by cleaving the R346-M347 bait peptide bond in the reactive center loop. No cleavage of PAI-1 was detected with Yersinia pseudotuberculosis, an oral/fecal pathogen from which Y. pestis has evolved, or with Escherichia coli. The cleavage and inactivation of PAI-1 were mediated by the outer membrane proteases plasminogen activator Pla of Y. pestis and PgtE protease of S. enterica, which belong to the omptin family of transmembrane endopeptidases identified in Gram-negative bacteria. Cleavage of PAI-1 was also detected with the omptins Epo of Erwinia pyrifoliae and Kop of Klebsiella pneumoniae, which both belong to the same omptin subfamily as Pla and PgtE, whereas no cleavage of PAI-1 was detected with omptins of Shigella flexneri or E. coli or the Yersinia chromosomal omptins, which belong to other omptin subfamilies. The results reveal a novel serpinolytic mechanism by which enterobacterial species expressing omptins of the Pla subfamily bypass normal control of host proteolysis.Plasminogen activator inhibitor 1 (PAI-1) is a key regulator of the mammalian fibrinolytic/plasminogen system (29, 37). The fibrinolytic system comprises the serine protease zymogen plasminogen, urokinase-type plasminogen activator (uPA), tissue-type plasminogen activator (tPA), PAI-1, and plasmin inhibitor ␣ 2 -antiplasmin (␣ 2 AP) (for a review, see reference 52). Plasminogen is converted to plasmin, which is a broad-spectrum serine protease that dissolves fibrin in blood clots, degrades laminin of basement membranes, and activates matrix metalloproteinases that degrade collagens and gelatins in tissue barriers. Herewith, plasmin controls physiological processes such as fibrinolysis/coagulation, cell migration and invasion, and tumor metastasis (29, 37). PAI-1 maintains normal hemostasis by inhibiting the function of the plasminogen activators tPA and uPA, which are serine proteases and highly specific for cleavage of the plasminogen molecule. tPA binds to fibrin and is associated with plasmin-mediated breakdown of fibrin clots, whereas uPA has low affinity for fibrin and associates with cell surface proteolysis, cellular migration, and damage of tissue barriers (52).The mammalian fibrinolytic and coagulation systems are targeted by invasive bacterial pathogens during infection (reviewed in references 6, 11, 34, and 61). In bacterial sepsis, increased production of fibrin clots at a damaged endothelium results from enhanced thrombin-catalyzed fibrin generation and from an increased serum level of PAI-1. Coagulation can protect the host by activating immune systems or by physically restraining the bacteria (6,15,25,41). On the other hand, several invasive bacterial pathogens enhance fibrinolysis eithe...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The Omptins ofYersinia pestisandSalmonella entericaCleave the Reactive Center Loop of Plasminogen Activator Inhibitor 1

et al. 2010

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“…aeruginosa clinical isolates derived from patients with infective endocarditis subsequent to implantation of a pacemaker [29]. Analysis of proteinases in bacterial secretomes by gelatin zymography and Elastin-Congo red hydrolysis assay was performed as reported [21,29,32,33], and is detailed in Protocol S1 in File S1. Experimental dilutions of secretomes or LB control medium are expressed as the percentage of bacterial culture supernatant in the final reaction volume.…”

Section: Methodsmentioning

confidence: 99%

Disruption of the Endothelial Barrier by Proteases from the Bacterial Pathogen Pseudomonas aeruginosa: Implication of Matrilysis and Receptor Cleavage

et al. 2013

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Within the vasculature, uncontrolled pericellular proteolysis can lead to disruption of cell-to-cell and cell-to-matrix interactions and subsequent detachment-induced cell apoptosis, or anoikis, contributing to inflammatory vascular diseases, with the endothelium as the major target. Most studies so far have focused on endogenous proteinases. However, during bloodstream infections, bacterial proteinases may also trigger endothelial anoikis. We thus investigated the potential apoptotic activity of the proteinases secreted by the haematotropic opportunistic pathogen, Pseudomonas aeruginosa, and particularly its predominant metalloproteinase, LasB. For this, we used the secretome of the LasB-expressing pseudomonal strain, PAO1, and compared it with that from the isogenic, LasB-deficient strain (PAO1∆lasB), as well as with purified LasB. Secretomes were tested for apoptotic activity on cultured human endothelial cells derived from the umbilical vein or from the cerebral microvasculature. We found that the PAO1 secretome readily induced endothelial cell anoikis, as did secretomes of LasB-positive clinical pseudomonal isolates, while the PAO1∆lasB secretome had only a limited impact on endothelial adherence and viability. Notably, purified LasB reproduced most of the effects of the LasB-containing secretomes, and these were drastically reduced in the presence of the LasB-selective inhibitor, phosphoramidon. A precocious and extensive LasB-dependent degradation of several proteins associated with the endothelial extracellular matrix, fibronectin and von Willebrand factor, was observed by immunofluorescence and/or immunoblotting analysis of cell cultures. Moreover, the PAO1 secretome, but not that from PAO1∆lasB, specifically induced rapid endoproteolysis of two major interendothelial junction components, VE-cadherin and occludin, as well as of the anti-anoikis, integrin-associated urokinase receptor, uPAR. Taken as a prototype for exogenous haemorrhagic proteinases, pseudomonal LasB thus appears to induce endothelial anoikis not only via matrilysis, as observed for many pro-apoptotic proteinases, but also via cleavage of some essential cell-to-cell and cell-to-matrix adhesion receptors implicated in the maintenance of the endothelial barrier.

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“…Secreted proteases targeting human coagulation or fibrinolysis proteins have been described in other pathogenic bacteria as well, e.g., Pseudomonas aeruginosa, Bacillus anthracis , and Staphylococcus aureus (Beaufort et al, 2008, 2010; Chung et al, 2011). Two main features distinguish Pla from these extracellular proteases.…”

Section: Yersinia Infections and The Hemostatic Controlmentioning

confidence: 99%

Fibrinolytic and coagulative activities of Yersinia pestis

Korhonen¹,

Haiko²,

Laakkonen³

et al. 2013

Front. Cell. Infect. Microbiol.

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The outer membrane protease Pla belongs to the omptin protease family spread by horizontal gene transfer into Gram-negative bacteria that infect animals or plants. Pla has adapted to support the life style of the plague bacterium Yersinia pestis. Pla has a β-barrel fold with 10 membrane-spanning β strands and five surface loops, and the barrel surface contains bound lipopolysaccharide (LPS) that is critical for the conformation and the activity of Pla. The biological activity of Pla is influenced by the structure of the surface loops around the active site groove and by temperature-induced LPS modifications. Several of the putative virulence-related functions documented for Pla in vitro address control of the human hemostatic system, i.e., coagulation and fibrinolysis. Pla activates human plasminogen to the serine protease plasmin and activates the physiological plasminogen activator urokinase. Pla also inactivates the protease inhibitors alpha-2-antiplasmin and plasminogen activator inhibitor 1 (PAI-1) and prevents the activation of thrombin-activatable fibrinolysis inhibitor (TAFI). These functions enhance uncontrolled fibrinolysis which is thought to improve Y. pestis dissemination and survival in the mammalian host, and lowered fibrin(ogen) deposition has indeed been observed in mice infected with Pla-positive Y. pestis. However, Pla also inactivates an anticoagulant, the tissue factor (TF) pathway inhibitor, which should increase fibrin formation and clotting. Thus, Pla and Y. pestis have complex interactions with the hemostatic system. Y. pestis modifies its LPS upon transfer to the mammalian host and we hypothesize that the contrasting biological activities of Pla in coagulation and fibrinolysis are influenced by LPS changes during infection.

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Activation of human pro-urokinase by unrelated proteases secreted by Pseudomonas aeruginosa

Cited by 18 publications

References 51 publications

The Omptins ofYersinia pestisandSalmonella entericaCleave the Reactive Center Loop of Plasminogen Activator Inhibitor 1

The Omptins ofYersinia pestisandSalmonella entericaCleave the Reactive Center Loop of Plasminogen Activator Inhibitor 1

Disruption of the Endothelial Barrier by Proteases from the Bacterial Pathogen Pseudomonas aeruginosa: Implication of Matrilysis and Receptor Cleavage

Fibrinolytic and coagulative activities of Yersinia pestis

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