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.

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.

Preparation and Properties of Human Prothrombin Complex

1970 ◽  
Vol 24 (03/04) ◽  
pp. 325-333 ◽  
Author(s):  
G. H Tishkoff ◽  
L. C Williams ◽  
D. M Brown
Keyword(s):  
Molecular Weight ◽  
Proteolytic Enzyme ◽  
Enzyme Inhibitor ◽  
Specific Activity ◽  
Crystalline Form ◽  
Sedimentation Equilibrium ◽  
Crystalline Complex ◽  
Interaction Product ◽  
Proteolytic Enzyme Inhibitor ◽  
Purification Scheme

SummaryAs a corollary to our previous studies with bovine prothrombin, we have initiated a study of human prothrombin complex. This product has been isolated in crystalline form as a barium glycoprotein interaction product. Product yields were reduced compared to bovine product due to the increased solubility of the barium glycoprotein interaction product. On occasion the crystalline complex exhibited good yields. The specific activity of the crystalline complex was 1851 Iowa u/mg. Further purification of human prothrombin complex was made by removal of barium and by chromatography on Sephadex G-100 gels. The final product evidenced multiple procoagulant activities (II, VII, IX and X). The monomeric molecular weight determined by sedimentation equilibrium in a solvent of 6 M guanidine-HCl and 0.5% mercaptoethanol was 70,191 ± 3,057 and was homogeneous with respect to molecular weight. This product was characterized in regard to physical constants and chemical composition. In general, the molecular properties of human prothrombin complex are very similar to the comparable bovine product. In some preparations a reversible proteolytic enzyme inhibitor (p-aminophenylarsonic acid) was employed in the ultrafiltration step of the purification scheme to inhibit protein degradation.

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.

scholarly journals Kinetic properties of the primary inhibitor of plasmin from human plasma

1977 ◽  
Vol 163 (2) ◽  
pp. 389-391 ◽  
Author(s):  
U Christensen ◽  
I Clemmensen
Keyword(s):  
Human Plasma ◽  
Enzyme Inhibitor ◽  
Kinetic Properties ◽  
Inhibitor Complex ◽  
Rapid Formation ◽  
Plasmin Inhibitor

The interaction of human plasmin with the newly discovered alpha2-plasmin inhibitor was investigated. It was found from rate measurements that the reaction involves the rapid formation of a first enzyme-inhibitor complex, followed by the slow irreversible transition to another complex. L-Lysine influences the first step, but not the second.

scholarly journals Lundep, a Sand Fly Salivary Endonuclease Increases Leishmania Parasite Survival in Neutrophils and Inhibits XIIa Contact Activation in Human Plasma

2014 ◽  
Vol 10 (2) ◽  
pp. e1003923 ◽  
Author(s):  
Andrezza C. Chagas ◽  
Fabiano Oliveira ◽  
Alain Debrabant ◽  
Jesus G. Valenzuela ◽  
José M. C. Ribeiro ◽  
...  
Keyword(s):  
Human Plasma ◽  
Sand Fly ◽  
Leishmania Parasite ◽  
Contact Activation ◽  
Parasite Survival

Sinapine Esterase I. Characterization of Sinapine Esterase from Cotyledons of Raphanus sativus

1979 ◽  
Vol 34 (9-10) ◽  
pp. 715-720 ◽  
Author(s):  
Gerhild Nurmann ◽  
Dieter Strack
Keyword(s):  
Enzyme Activity ◽  
Esterase Activity ◽  
Raphanus Sativus ◽  
Sinapic Acid ◽  
Inhibitory Effect ◽  
Ph Optimum ◽  
Diisopropyl Fluorophosphate ◽  
E 600 ◽  
Red Radish

Abstract From cotyledons of Raphanus sativus (red radish) an esterase activity which catalyzes the hy­drolysis of sinapine into sinapic acid and choline has been isolated. The enzyme, which has a near absolute specificity, is not analogous with any esterase described in the literature. The reaction has a pH optimum of 8.5 and the apparent Km is 1.95 × 10-5 m. The enzyme is relatively insensi­tive to both physostigmine (eserine) {Ki = 1.73 × 10-4 m) and neostigmine (Ki = 2 .1 3 × 10-4 ᴍ). Diisopropyl fluorophosphate (DFP) showed no inhibition and diethyl p-nitrophenylphosphate (E 600) only a slight inhibitory effect at 10-5 ᴍ, respectively. Choline (10-2 ᴍ) was inhibitory but acetylcholine (10-2 ᴍ) stimulated the enzyme activity.

The activation of human plasma prekallikrein as a hemocompatibility test f or biomaterials. II. Contact activation by EVAL and EVAL-SMA copolymers

1993 ◽  
Vol 4 (3) ◽  
pp. 643-652 ◽  
Author(s):  
Giulio D. Guerra ◽  
Niccoletta Barbani ◽  
Luigi Lazzeri ◽  
Letizia Lelli ◽  
Maurizio Palla ◽  
...  
Keyword(s):  
Human Plasma ◽  
Contact Activation

scholarly journals Sulfatide bilayers as a surface for contact activation in human plasma

1987 ◽  
Vol 149 (3) ◽  
pp. 1002-1007 ◽  
Author(s):  
G. Tans ◽  
A.J. Verkleij ◽  
J Yu ◽  
J.H. Griffin
Keyword(s):  
Human Plasma ◽  
Contact Activation
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