scholarly journals Nonplasminogen-dependent protease in human plasma

Blood ◽  
1979 ◽  
Vol 54 (3) ◽  
pp. 607-613
Author(s):  
JA Marcum ◽  
DL Kline
Keyword(s):  
Ionic Strength ◽  
Human Plasma ◽  
Trypsin Inhibitor ◽  
Fibrinolytic Activity ◽  
Coagulation Factors ◽  
Contact Activation ◽  
Ph Optimum ◽  
Salt Addition ◽  
Caseinolytic Activity ◽  
Pancreatic Trypsin Inhibitor

Abstract Equal volumes of plasma and 0.3 M K2HPO4, pH 7.4, were mixed, diluted 20-fold, and adjusted to pH 5.2. After incubation at 37 degrees C for 30 min, the euglobulin percipitate, redissolved in 0.1 M K2HPO4, pH 7.4, developed caseinolytic activity (0.05 CTA U/ml). Na2HPO4 or NaCl of similar ionic strength could replace K2HPO4. The pH optimum of the protease was 6.5, activity falling off sharply below pH 6.0 and above 7.4. The proteolytic activity was inhibited by diisopropylphosphofluoridate and by pancreatic trypsin inhibitor, but was not inhibited by soybean trypsin inhibitor. The activity was not due to plasmin, contact activation, or coagulation factors, since it was fully generated in plasminogen-depleted, factors XII, XI, VII deficient, and prekallikrein-deficient plasmas. Purified Cl-esterase was not caseinolytic in our system. Redissolved euglobulin precipitate prepared from normal plasma without salt addition could serve as starting material for the generation of caseinolytic activity, as could serum, indicating that the Hageman factor cofactor and thrombin are not required. The protease had no detectable procoagulant or fibrinolytic activity.

scholarly journals Nonplasminogen-dependent protease in human plasma

Blood ◽  
1979 ◽  
Vol 54 (3) ◽  
pp. 607-613
Author(s):  
JA Marcum ◽  
DL Kline
Keyword(s):  
Ionic Strength ◽  
Human Plasma ◽  
Trypsin Inhibitor ◽  
Fibrinolytic Activity ◽  
Coagulation Factors ◽  
Contact Activation ◽  
Ph Optimum ◽  
Salt Addition ◽  
Caseinolytic Activity ◽  
Pancreatic Trypsin Inhibitor

Equal volumes of plasma and 0.3 M K2HPO4, pH 7.4, were mixed, diluted 20-fold, and adjusted to pH 5.2. After incubation at 37 degrees C for 30 min, the euglobulin percipitate, redissolved in 0.1 M K2HPO4, pH 7.4, developed caseinolytic activity (0.05 CTA U/ml). Na2HPO4 or NaCl of similar ionic strength could replace K2HPO4. The pH optimum of the protease was 6.5, activity falling off sharply below pH 6.0 and above 7.4. The proteolytic activity was inhibited by diisopropylphosphofluoridate and by pancreatic trypsin inhibitor, but was not inhibited by soybean trypsin inhibitor. The activity was not due to plasmin, contact activation, or coagulation factors, since it was fully generated in plasminogen-depleted, factors XII, XI, VII deficient, and prekallikrein-deficient plasmas. Purified Cl-esterase was not caseinolytic in our system. Redissolved euglobulin precipitate prepared from normal plasma without salt addition could serve as starting material for the generation of caseinolytic activity, as could serum, indicating that the Hageman factor cofactor and thrombin are not required. The protease had no detectable procoagulant or fibrinolytic activity.

AUTOACTIVATION OF HUMAN PLASMA PREKALLIKREIN

1987 ◽  
Author(s):  
G Tans ◽  
J Rosing ◽  
M Berrettini ◽  
B Lammle ◽  
J H Griffin
Keyword(s):  
Ionic Strength ◽  
Human Plasma ◽  
Rate Constant ◽  
Trypsin Inhibitor ◽  
Active Form ◽  
Second Order ◽  
Reversible Inhibitor ◽  
Lag Phase ◽  
Contact Activation ◽  
Amidolytic Activity

Incubation of purified human plasma prekallikrein with sulfatides or dextran sulfate resulted in spontaneous activation of prekallikrein as judged by the appearance of amidolytic activity towards the chromogenic substrate H-D-pro-phe-arg-p-nitroanilide (S 2302). The time course of generation of amidolytic activity was sigmoidal with an apparent lag phase followed by a rapid activation until finally a plateau was reached. Soybean trypsin inhibitor completely blocked prekallikrein activation whereas corn, limabean and ovomucoid trypsin inhibitor did not. The Ki of the reversible inhibitor, benzamidine, for autoactivation (240 uM) was identical to the Ki of benzamidine for kallikrein. Thus, spontaneous prekallikrein activation and kallikrein showed the same specificity for a number of serine protease inhibitors, indicating that prekallikrein is activated by its own enzymatically active form, kallikrein. Immunoblotting analysis showed that, concomitant with the appearance of amidolytic activity, prekallikrein was cleaved. However, prekallikrein was not quantitatively converted into two-chain kallikrein since other polypeptide products were visible on the gels. This accounts for the observation that in amidolytic assays not all prekallikrein present in the reaction mixture was measured as active kallikrein. Kinetic analysis showed that prekallikrein activation can be described by a second-order reaction mechanism in which prekallikrein is activated kallikrein. The apparent second order rate constant was 27000 M-ls-1 (pH 7.2, 50 uM sulfatides, ionic strength 1=0.06, at 37°C). Autocatalytic prekallikrein activation was strongly dependent on the ionic strength, since there was a considerable decrease in the rate of the reaction at high salt concentrations. Our data support a prekallikrein autoactivation mechanism in which surface-bound kallikrein activates surface-bound prekallikrein. The rate constant of autoactivation is considerably lower than the rate constants reported for Factor Xlla dependent prekallikrein formation. Autocatalytic prekallikrein activation may, however, contribute to kallikrein formation during the initiating phase of contact activation.

Differentiation of Trypsin-Like Enzymes in Human Plasma

1972 ◽  
Vol 18 (11) ◽  
pp. 1385-1394 ◽  
Author(s):  
Herbert D Gullick
Keyword(s):  
Human Plasma ◽  
Esterase Activity ◽  
Enzyme Inhibitor ◽  
Heat Stability ◽  
Differential Sensitivity ◽  
Contact Activation ◽  
Ph Optimum ◽  
Proteolytic Enzyme Inhibitor ◽  
Optimum Heat ◽  
Stability Effect

Abstract The arginine amidase and arginine esterase activity of human plasma is compared with that of trypsin, thrombin, plasmin, and kallikrein by using the homologous synthetic amino acid substrates, benzoyl arginine amide (BAA) and benzoyl arginine ethyl ester (BAEE). Hydrolyses of BAA and BAEE in plasma are distinguished by several features: pH optimum, heat stability, storage stability, effect of dilution, surface contact activation, streptokinase activation, inhibition by proteolytic enzyme inhibitor, and range of normal variation. The differential sensitivity of trypsin, thrombin, plasmin, and kallikrein toward these substrates and the similarity in reaction characteristics of plasma-BAA and trypsin-BAA provide evidence that the arginine amidase activity of plasma represents trypsin, while the arginine esterase activity of plasma is nonspecific.

On the Combined Administration of Streptokinase and Pancreatic Trypsin Inhibitor

1964 ◽  
Vol 12 (02) ◽  
pp. 391-395 ◽  
Author(s):  
M Verstraete ◽  
J Vermylen ◽  
C Vermylen ◽  
R. A De Vreker
Keyword(s):  
Trypsin Inhibitor ◽  
Fibrinolytic Activity ◽  
Fibrinogen Level ◽  
Preliminary Results ◽  
Combined Administration ◽  
Pancreatic Trypsin Inhibitor

SummaryPreliminary results on the modification of blood fibrinogen level and plasma plasminogen value and fibrinolytic activity during combined administration of streptokinase and pancreatic trypsin inhibitor are presented. Not only fibrinogen fall but also plasminogen depletion is inhibited.

Proteolytic Activity of the Activator Produced by Streptokinase in Human Plasma

1971 ◽  
Vol 26 (01) ◽  
pp. 088-098
Author(s):  
H Claeys ◽  
A Amery ◽  
R Verhaeghe
Keyword(s):  
Human Plasma ◽  
Plasminogen Activator ◽  
Proteolytic Activity ◽  
Trypsin Inhibitor ◽  
Soybean Trypsin Inhibitor ◽  
Plasmin Activity ◽  
In Vitro Activation ◽  
Caseinolytic Activity ◽  
Fibrinogenolytic Activity

SummaryIn the in vitro activation of the fibrinolytic system of human plasma by streptokinase (SK) maximal caseinolytic and fibrinogenolytic activity was found at a SK concentration of 2.103 u/ml plasma. At higher concentrations of SK these activities decrease and level off to a plateau at a SK concentration of 105 u/ml plasma for the caseinolytic activity and 104 u/ml plasma for the fibrinogenolytic activity. At these high SK concentrations the caseinolytic and fibrinogenolytic activity can not or only slightly be inhibited by soybean trypsin inhibitor at concentrations, which completely inhibit these activities at lower SK concentrations. The bovine plasminogen activator activity increases with increasing SK concentrations between 103 and 106 u SK/ml plasma. No plateau of maximal activator activity could be demonstrated. It is suggested that the remaining fibrinogenolytic and caseinolytic activity at high SK concentrations is not due to plasmin activity, but that the bovine plasminogen activator has direct caseinolytic and fibrinogenolytic activity.

Dextran Sulfate Stimulated Fibrinolytic Activity In Whole Human Plasma: Dependence On The Contact Activation System And On 4 Urokinase-Related Antigen

1981 ◽  
Author(s):  
Lindsey A Miles ◽  
Zuleika Rothschild ◽  
John H Griffin
Keyword(s):  
Human Plasma ◽  
Fibrinolytic Activity ◽  
Dextran Sulfate ◽  
Factor Xii ◽  
Dependent Manner ◽  
Contact Activation ◽  
Prolonged Incubation ◽  
Normal Plasma ◽  
Fibrin Plate ◽  
Activation System

The generation of fibrinolytic activity in whole human plasma in the presence of dextran sulfate was studied. Plasma was preincubated with N-flufenamyl-β-alanine to remove its antiplasmin and anti activator activities and then incubated with 50 μg/ml dextran sulfate (Mr - 500,000) for 30 min at 40. The initial fibrinolytic activity in 30 min, as assessed on a 125I-fibrin plate, was equivalent to approximately 9 ng/ml purified plasmin. A fraction of goat antibodies to plasminogen blocked the fibrinolytic activity of stimulated plasma, indicating that the activity was plasminogen dependent. Plasmas genetically deficient in either prekallikrein or Factor XII (Hageman Factor) showed a diminished initial rate of generation of fibrinolytic activity in response to dextran sulfate. However, after prolonged incubation (∼3 hr) of stimulated deficient plasmas with fibrin, the fibrinolytic activity approached that of stimulated normal plasma. When normal plasma was preincubated with the gamma fraction of goat antibodies made against purified urokinase, the dextran sulfate stimulated fibrinolytic activity was markedly decreased in a dose dependent manner. The data suggest that the fibrinolytic activity stimulated in whole human plasma in the presence of dextran sulfate and N-flufenamyl-β-alanine is dependent upon proteins of the contact activation system and also upon molecules immunologically related to urokinase.

The Adsorption of Blood Coagulation Factors II, VII, IX and X from Human Plasma to Aluminium Hydroxide

1972 ◽  
Vol 27 (03) ◽  
pp. 490-501 ◽  
Author(s):  
A. C. W Swart ◽  
B. H. M Klaassen ◽  
C. H. F Bloys-van Treslong-De Groot ◽  
H. C Hemker
Keyword(s):  
Plasma Concentration ◽  
Ionic Strength ◽  
Human Plasma ◽  
Blood Coagulation ◽  
Aluminium Hydroxide ◽  
Coagulation Factors ◽  
Blood Coagulation Factors ◽  
Adsorption Time ◽  
Normal Human ◽  
Influence Of Ph

SummaryThe influence of pH, adsorption time, ionic strength, buffer type and plasma concentration on adsorption and elution of the factors of the prothrombin complex from normal human ACD plasma onto A1(OH)3 is investigated. Reproducible small differences are found between the behaviour of the four factors II, VII, IX, and X.

Inactivation of Antiplasmin and Complement C1 in Human Plasma Rendered Fibrinolytic by Synthetic Organic Compounds

1963 ◽  
Vol 10 (01) ◽  
pp. 151-163 ◽  
Author(s):  
Kurt N von Kaulla
Keyword(s):  
Human Plasma ◽  
Organic Compounds ◽  
Bovine Serum ◽  
Fibrinolytic Activity ◽  
Plasma Generation ◽  
Synthetic Organic Compounds

SummaryCertain synthetic organic compounds induce upon dissolution marked fibrinolytic activity in human plasma, reduce the antiplasmin titer of human or bovine serum and destroy the complement C1 of human plasma. Generation of fibrinolytic activity and reduction of antiplasmin are concentration-depending time reactions. Destruction of complement C1 occurs almost instantaneously. Minor molecular modifications abolish all three activities of the compounds.

Inhibition of Coagulation and Fibrinolysis by Aromatic Amidino Compounds

1971 ◽  
Vol 25 (03) ◽  
pp. 391-404 ◽  
Author(s):  
J.D Geratz
Keyword(s):  
Prothrombin Time ◽  
Human Plasma ◽  
Partial Thromboplastin Time ◽  
Inhibitory Effect ◽  
Clot Lysis ◽  
Similar Degree ◽  
Contact Activation ◽  
Plasma Clot ◽  
Human Thrombin ◽  
Canine Plasma

Summary1. Aromatic diamidines which are potent inhibitors of trypsin possess a marked inhibitory effect on the clotting activity of human thrombin and on the prothrombin time and partial thromboplastin time of human plasma. They also block the contact activation phase of the coagulation process. The strongest inhibitor among the compounds tested was M & B 4596 which was followed in second place by pentamidine.2. Pentamidine was 10 times more active than ε-ACA in impeding streptokinase-induced lysis of human plasma clots. It was 100-200 times stronger than ε-ACA in inhibiting the activation of bovine plasminogen by activators formed from the interaction between streptokinase and either human plasmin(ogen) or human plasma.3. The prothrombin time and partial thromboplastin time of canine plasma were less susceptible to inhibition by pentamidine than the same tests on human plasma. Clot lysis in the canine system was inhibited by pentamidine to a similar degree as in the human system. After intravenous injection of pentamidine in the dog there occurred the expected prolongation of the partial thromboplastin time and of the clot lysis time.

Interaction of Heparin with Fibrinolysis

1963 ◽  
Vol 09 (02) ◽  
pp. 446-458 ◽  
Author(s):  
Rudolf Holemans ◽  
Dionysios Adamis ◽  
James F Horace
Keyword(s):  
Human Plasma ◽  
Fibrinolytic Activity ◽  
Plasminogen Activation ◽  
Fibrinolytic Agents ◽  
High Concentration ◽  
Enhancing Effect

SummaryHeparin in high concentration inhibits the fibrinolysis of human plasma clots or bovine fibrin by fibrinolytic agents which produce plasminogen activation. Heparin has no effect on the fibrinolytic activity of plasmin or Aspergillus protease.In order to produce inhibition of plasminogen activation heparin requires the presence of a co-factor which is present in citrated human plasma but absent from its euglobulin fraction.In none of the concentrations tested has heparin an enhancing effect on fibrinolysis.

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