• BMB Reports
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• v.37 no.2
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• pp.199-205
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• 2004

The six lumbrokinase fractions (F1 to F6) with fibrinolytic activities were purified from earthworm Lumbricus rubellus lysates using the procedures of autolysis, ammonium sulfate fractionation, and column chromatography. The proteolytic activities on the casein substrate of the six iso-enzymes ranged from 11.3 to 167.5 unit/mg with the rank activity orders of F2 > F1 > F5 > F6 > F3 > F4. The fibrinolytic activities of the six fractions on the fibrin plates ranged from 20.8 to 207.2 unit/mg with rank orders of F6 > F2 > F5 > F3 > F1 > F4. The molecular weights of each iso-enzyme, as estimated by SDS-PAGE, were 24.6 (F1), 26.8 (F2), 28.2 (F3), 25.4 (F4), 33.1 (F5), and 33.0 kDa (F6), respectively. The plasminogen was activated into plasmin by the enzymes. The optimal temperature of the six iso-enzymes was $50^{\circ}C$, and the optimal pH ranged from pH 4-12. The four iso-enzymes (F1-F4) were completely inhibited by PMSF. The two enzymes (F5 and F6) were completely inhibited by aprotinin, TLCK, TPCK, SBTI, LBTI, and leupeptin. The N-terminal amino acid (aa) sequences of the first 20 to 22 residues of each fraction had high homology. All six isoenzymes had identical aa residues 2-3 and 13-15. The N-terminal 21-22 aa sequences of the F2, F3, and F4 isoenzymes were almost the same. The N-terminal aa sequences of F5 and F6 were identical.

• Tuberculosis and Respiratory Diseases
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• v.40 no.4
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• pp.378-383
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• 1993

Background: In recent reports, it has been reported that increased coagulation and decreased fibrinolytic activity has been responsible for abnormal fibrin turnover in exudative pleural effusion. In the cases of empyema, the fibrinopurulent stage is characterized by the fibrin deposition resulting in formation of limiting membranes in the visceral and parietal pleura. Recently attention has been focused on intrapleural fibrinolytic therapy capable of removing intrapleural fibrin deposits by urokinase (UK) in the treatment of empyema. However, these clinical trials have provided the clinical evidences for resolution of pleural loculation after intrapleural urokinase injection (UK-injection), the profiles of fibrinolytic activity following the treatment were still not investigated. Therefore in order to demonstrate the fibrinolytic evidences behind the clinical efficacy of intracavitary UK-injection, we examined intrapleural plasminogen activator activity (PA-activity) and D-dimer (D-Di) concentrations before and after each repeated UK-injection into the pleura in subjects with loculated empyema cavity. Methods: In a group of 14 patients with multiple loculated empyema cavity, PA-activity and D-Di concentrations were measured before and after repeated UK-injection. One hundred thousand IU of UK was injected at each time and all sujects had at least two times of UK injection accoring to clinical decisions. Nine out of 14 sujects had three times of UK-injection. Results: The mean (${\pm}SE$) PA-activity prior to treatemnt was $10.5{\pm}7.0$ and it was increased to $91.9{\pm}27.0,\;432.3{\pm}177.1,\;170.0{\pm}85.3$ IU tPA/ml after first, second and third time of UK-injection respectively (p<0.01). D-Di concentrations were also increased from $4.16{\pm}1.06{\times}10^5$ to $9.62{\pm}1.54{\times}10^5,\;12.31{\pm}1.89{\times}10^5,\;8.54{\pm}1.56{\times}10^5$ ng/ml in the same order as above (p<0.05). Conclusion: The suppressed fibrinolytic activity in the empyema cavity get removed sinificantly after inrracavitary injection of urokinase by generation of additional intrapleural plasmin.