Pesticins III. Expression of Coagulase and Mechanism of Fibrinolysis

Abstract

Mutational loss of pesticin I, a bacteriocin-like substance produced by Pasteurella pestis, is known to result in concomitant loss of a coagulase and fibrinolytic factor. No relationship was detected between pesticinogeny and other tested properties either associated with virulence or peculiar to P. pestis. Pesticin I was distinguished from the coagulase and fibrinolytic activities on the basis of anatomical distribution, behavior during gel filtration, and sensitivity to heat. Coagulase and the fibrinolytic factor were not differentiated by these criteria. Spontaneous suppressor mutations causing reversion to pesticinogeny were not detected, nor were such mutants obtained by treatment with ultraviolet light or 2-aminopurine. Attempts to demonstrate a common activator of pesticin I, coagulase, or the fibrinolytic factor in extracts of pesticinogenic cells were not successful. These results are in accord with the hypothesis that at least two structural genes for the three activities reside on a replicon distinct from the chromosome proper. Fibrinolytic activity was significantly reduced in the presence of 0.003 M e-aminocaproic acid and was nonexistent on fibrin films freed from endogenous plasminogen by treatment with heat. Fibrinolytic activity on heated films could be restored by addition of plasma or serum from six mammalian species. Accordingly, the plague fibrinolytic factor, like staphylokinase or urokinase, promotes the conversion of plasminogen to plasmin.

Immune Response to Yersinia Outer Proteins and Other Yersinia pestis Antigens after Experimental Plague Infection in Mice

“…This result supports the continued use of detection of antibodies to F1 for serodiagnosis of human plague (46). Both F1, a capsular protein, and Pla, a membrane protein known to be on the surface of the bacteria (5,52), should be readily accessible to the immune system of the host. Consequently, it is not surprising that after combining data for all of the antibiotic treatment groups, we found that the highest percentages of animals responded to F1 and Pla (95 and 62%, respectively).…”

A Yersinia pestis‐specific DNA fragment encodes temperature‐dependent coagulase and fibrinolysin associated phenotypes

“…For example, expression of the coagulase phenotype may require the Pla protein to be inserted in the outer membrane, while the fibrinolytic form of the protein could be active in an aqueous environment in the periptasm or supernatant. Such a model could explain why the coagulase phenotype is easily observed with whole cells but is difficult to obtain in cell extracts (Beesley et aL, 1967; this study), while fibrinolytic activity follows the opposite pattern. Further biochemical analysis will be required to answer these questions satisfactorily.…”

“…It is clear from these data and the work of Sodeinde and Goguen (1988) that a single gene {pla) Is responsible for both coagulase and fibrinolytic activities. Any attempts to explain how this occurs must take into account the following; (i) the modulation between coagulase and fibrinolytic activities is temperature-dependent, occurs post-translationally and appears to be reversible; (ii) the probable target for the fibrinolytic activity is plasminogen rather than fibrin (Beesley et al, 1967); (iii) the Pla factor has been associated with additional proteolytic activities ; and (iv) two forms of the pla gene product have been described (Sodeinde and Goguen, 1988;Sample et al. 1987;Straley and Brubaker, 1982).…”

“…Fibrin films were generated according to the methods of Beesley et al (1967) with modifications. Briefly, 10ml of 0.5% fibrinogen (bovine, 90% clottable, or human, 65% ciottable, from United States Biochemicals Corp., Cleveland, OH) were placed in a 75mm petri dish and clotted with the addition of 25 NIH Units of bovine thrombin (USB) in borate buffer (Lewis and Ferguson, 1950) at a concentration of 1 Unit \x.\ '.…”

Plasmids in Yersinia pestis

“…Yersiniae of diverse origin and their isogenic mutants lacking selected determinants of virulence were characterized as previously described (1). The organisms were aerated at 26°C (100 ml of Difco heart infusion broth per 1-liter Erlenmeyer flask) on a shaker and harvested by centrifugation (10,000 x g for 10 min).…”