Expression of plasminogen activator pla of Yersinia pestis enhances bacterial attachment to the mammalian extracellular matrix - PubMed (original) (raw)

Expression of plasminogen activator pla of Yersinia pestis enhances bacterial attachment to the mammalian extracellular matrix

K Lähteenmäki et al. Infect Immun. 1998 Dec.

Abstract

The effect of the plasminogen activator Pla of Yersinia pestis on the adhesiveness of bacteria to the mammalian extracellular matrix was determined. Y. pestis KIM D27 harbors the 9.5-kb plasmid pPCP1, encoding Pla and pesticin; the strain efficiently adhered to the reconstituted basement membrane preparation Matrigel, to the extracellular matrix prepared from human lung NCI-H292 epithelial cells, as well as to immobilized laminin. The isogenic strain Y. pestis KIM D34 lacking pPCP1 exhibited lower adhesiveness to both matrix preparations and to laminin. Both strains showed weak adherence to type I, IV, and V collagens as well as to human plasma and cellular fibronectin. The Pla-expressing recombinant Escherichia coli LE392(pC4006) exhibited specific adhesiveness to both extracellular matrix preparations as well as to laminin. The Pla-expressing strains showed a low-affinity adherence to another basement membrane component, heparan sulfate proteoglycan, but not to chondroitin sulfate proteoglycan. The degradation of radiolabeled laminin, heparan sulfate proteoglycan, or human lung extracellular matrix by the Pla-expressing recombinant E. coli required the presence of plasminogen, and degradation was inhibited by the plasmin inhibitors aprotinin and alpha2-antiplasmin. Our results indicate a function of Pla in enhancing bacterial adhesion to extracellular matrices. Y. pestis also exhibits a low level of Pla-independent adhesiveness to extracellular matrices.

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Figures

FIG. 1

FIG. 1

Adherence of Pla+ Y. pestis KIM D27, Pla− Y. pestis KIM D34, Pla+ E. coli LE392(pC4006), and Pla− E. coli LE392(pC4007) to the reconstituted mouse BM preparation Matrigel and to the ECM prepared by detergent extraction from human lung NCI-H292 epithelial cells. The control surface was coated with BSA. The bacteria were tested at a concentration of 109 cells/ml.

FIG. 2

FIG. 2

Adherence of Pla+ Y. pestis KIM D27 and E. coli LE392(pC4006) as well as of Pla− Y. pestis KIM D34, E. coli LE392(pC4007), and E. coli LE392 to the BM preparation Matrigel reconstituted on glass slides as well as to the ECM prepared by detergent extraction from human lung NCI-H292 cells. The bacteria were tested at four different concentrations as indicated, and the data shown are means ± SDs for 20 randomly chosen microscopic fields of 1.6 × 104 μm2. The control surface was coated with BSA.

FIG. 3

FIG. 3

Adherence of Pla+ Y. pestis KIM D27 and E. coli LE392(pC4006) as well as of Pla− Y. pestis KIM D34, E. coli LE392(pC4007), and E. coli LE392 to immobilized proteins of the ECM. Bacterial concentration varied from 108 to 5 × 109 cells/ml, and the surface concentration of the target proteins was 2.5 pmol. The control surfaces were coated with the highly glycosylated fetuin and with the nonglycosylated BSA from a solution of 25 μg/ml. Data are means ± SDs for 20 randomly chosen microscopic fields of 1.6 × 104 μm2.

FIG. 4

FIG. 4

Adherence of Pla+ Y. pestis KIMD27 and E. coli LE392(pC4006) as well as of Pla− Y. pestis KIM D34, E. coli LE392(pC4007), and E. coli LE392 to immobilized proteoglycans. For comparison, bacterial adhesiveness to laminin and BSA is also shown. All target proteins were coated from a solution of 50 μg/ml, and the bacteria were tested at the concentrations of 108 cells/ml (A) and 109 cells/ml (B). Data are means ± SDs for 20 randomly chosen fields of 1.6 × 104 μm2.

FIG. 5

FIG. 5

Degradation of 125I-labeled laminin (A) and 35S-labeled extracellular matrix prepared from human lung NCI-H292 cells (B) by Pla+ or Pla− recombinant E. coli in the presence of plasminogen and inhibitors of plasmin activity. (C) Plasmin activity associated with the E. coli cells or in PBS without added bacteria, measured with the chromogenic substrate. Bacteria were used at a concentration of 2 × 109 cells/ml.

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