Studies on the Activation Mechanism of Fibrinolytic Enzyme System in Plasma by Human Pancreatic Elastase

1982 ◽  
Vol 47 (01) ◽  
pp. 008-013 ◽  
Author(s):  
N Toki ◽  
S Takasugi ◽  
K Ishizu ◽  
H Sato ◽  
H Sumi
Keyword(s):  
Molecular Weight ◽  
Enzyme System ◽  
Fibrinolytic Enzyme ◽  
Activation Mechanism ◽  
Low Molecular Weight ◽  
Pancreatic Elastase ◽  
Α2 Macroglobulin ◽  
Plasmin Inhibitor

SummaryIn the present study, the activation mechanism of fibrinolytic enzyme system in plasma by human pancreatic elastase was investigated. It was confirmed that human pancreatic elastase not only converted the co-existing plasminogen to low molecular weight-plasminogen which could be easily activated by the activator, but also inhibited α2-macroglobulin and α2-plasmin inhibitor which are antiactivators or fast reacting antiplasmins, and consequently, induced the activation of the fibrinolytic enzyme system in plasma.

α2–plasmin Inhibitor And α2-Macroglobulin-Plasmin Complexes In Plasma. Quantitation By An Enzyme-Linked Immunosorbent Assay

1981 ◽  
Author(s):  
P Harpel
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme System ◽  
Fold Increase ◽  
Fibrinolytic Enzyme ◽  
Enzyme Linked Immunosorbent Assay ◽  
Normal Individuals ◽  
Α2 Macroglobulin ◽  
Low Levels ◽  
Enzyme Complexes ◽  
Plasmin Inhibitor

An enzyme-linked immunosorbent assay has been developed for the quantitation of α2-plasmin inhibitor-plasmin and α2-macroglobulin-plasmin complexes. In this method, the inhibitor-plasmin complex is bound to a surface by an inhibitor specific antibody, then the plasmin bound to the inhibitor is quantified by a second antibody labeled with alkaline phosphatase. Inhibitor-enzyme complexes were generated in plasma by the addition of plasmin or of urokinase (UK). The concentration of plasmin added was well below the plasma concentration of α2-plasmin inhibitor or of α2-macroglobulin so that neither inhibitor would be fully saturated with enzyme. Under these conditions, increasing amounts of plasmin generated an increase in both α2-plasmin inhibitor-plasmin and α2-macroglobulin- plasmin complexes. Varying amounts of plasmin were incubated with each of the purified inhibitors and the complexes that formed were quantitated by immunoassay. These studies made it possible to quantitate the distribution of plasmin between the two inhibitors in plasma. In plasmin-treated plasma, 10% or less of the plasmin bound to both inhibitors was in complex with α2-macroglobulin. In contrast, between 19-51% of the plasmin generated in UK-activated plasma was bound to α2-macroglobulin. Thus major changes in the distribution of plasmin were observed depending upon whether plasmin was added to plasma or whether plasminogen was activated endogenously. 23 normal individuals had low levels of α2-plasmin inhibitor-plasmin complexes (4.1±3.5 fmol/ml) whereas six patients with laboratory evidence for DIC demonstrated a 16 to 35-fold increase in the concentration of these complexes. These data indicate that a useful new probe for the study of the fibrinolytic enzyme system has been developed.

The Kidney and Fibrinolysis

1970 ◽  
Vol 24 (01/02) ◽  
pp. 026-032 ◽  
Author(s):  
N. A Marsh
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Enzyme System ◽  
Normal Animal ◽  
Fibrinolytic Enzyme ◽  
The Other ◽  
Exclusion Chromatography ◽  
Normal Urine ◽  
Renal Origin ◽  
Molecular Exclusion Chromatography

SummaryMolecular exclusion chromatography was performed on samples of urine from normal and aminonucleoside nephrotic rats. Normal urine contained 2 peaks of urokinase activity, one having a molecular weight of 22,000 and the other around 200,000. Nephrotic urine contained three peaks of activity with MW’s 126,000, 60,000 and 30,000. Plasma activator determined from euglobulin precipitate had a MW. in excess of 200,000. The results indicate that in the normal animal, plasma plasminogen activator does not escape into the urine in substantial quantities but under the conditions of extreme proteinuria there may be some loss through the kidney. The alteration in urokinase output in nephrotic animals indicates a greatly disordered renal fibrinolytic enzyme system.The findings of this study largely support the hypothesis that plasma plasminogen activator of renal origin and urinary plasminogen activator (urokinase) are different molecular species.

A low molecular weight chymotrypsin-like novel fibrinolytic enzyme from Streptomyces sp. CS624

2011 ◽  
Vol 46 (7) ◽  
pp. 1449-1455 ◽  
Author(s):  
Poonam Mander ◽  
Seung Sik Cho ◽  
Jaya Ram Simkhada ◽  
Yun Hee Choi ◽  
Jin Cheol Yoo
Keyword(s):  
Molecular Weight ◽  
Fibrinolytic Enzyme ◽  
Low Molecular Weight ◽  
Streptomyces Sp

scholarly journals A Low Molecular Weight Fibrinolytic Enzyme from Polymorphonuclear Leukocytes (PMN)

1977 ◽  
Author(s):  
L.A. Moroz
Keyword(s):  
Molecular Weight ◽  
Solid Phase ◽  
Degradation Products ◽  
Gel Filtration ◽  
Protein Band ◽  
Fibrinolytic Enzyme ◽  
Low Molecular Weight ◽  
Cationic Protein ◽  
Alkaline Proteinase ◽  
Solid Phase Assay

Despite evidence implicating PMN in fibrinolysis, the enzymes involved are incompletely characterized.PMN were prepared from normal blood by dextran sedimentation and fibrinolytic activity assayed by l25I-fibrin solid phase assay (Blood 46:543, 1975). More than 80% of activity was associated with intact PMN, was stimulated by Na salicylate (+65%, 20 mg/100 ml) and inhibited by α1-anti-trypsin (α1AT, -48%, 2 × 10-6 M). Similar activity was found in a PMN membrane fraction prepared by homogenization, differential centrifugation and Sepharose 4B gel filtration, from which fraction it was released by freeze-thawing and/or IM KCl treatment. Soluble enzyme activity was inhibited by of α1AT (-80%, 10-6 M), PMSF (-98%, 10-3 M), FeCl3 and ZnCl3 (-100%, 10-2 M), vitamin E(-38%, 10-4 M) and trypan blue (-40%, 10-3 M), but not by EACA (10-2 M), tranexamic acid (10-2 M), TLCK (10-3 M) or TPCK (10-3 M). This activity had an alkaline proteinase pH-activity profile, was localized to a single cationic protein band on acid Polyacrylamide disc gel electrophoresis, with pl of 8.6-8.7 by isoelectric focussing, and eluted between lysozyme and myoglobin on Bio-Gel P-10. On Bio-Gel A 0.5m and P-10, 125I-fibrin degradation products eluted after myoglobin. These findings indicate the presence in PMN of a low molecular weight, membrane-associated fibrinolytic enzyme of alkaline proteinase and serine active site type.

In vivo activation of the fibrinolytic enzyme system of dogs

1963 ◽  
Vol 18 (2) ◽  
pp. 334-336 ◽  
Author(s):  
Fletcher B. Taylor ◽  
John Singleton ◽  
Arthur F. Bickford
Keyword(s):  
Blood Volume ◽  
Immune Reaction ◽  
Fibrinogen Level ◽  
Enzyme System ◽  
Fibrinolytic Enzyme ◽  
Plasma Fibrinogen ◽  
Clot Lysis ◽  
Blood Clots ◽  
Plasmin Inhibitor

We studied the effect of streptokinase and small amounts of human plasmin on the components of the fibrinolytic enzyme system of the dog. A 1-hr infusion of streptokinase (500 U/ml blood volume) and plasmin (.001 mg/ml blood volume) in dogs produced during the infusion: 1) No adverse immune reaction. 2) Enhanced lysis of blood clots. 3) Decreased serum plasminogen and plasma plasmin inhibitor levels. 4) Decreased plasma fibrinogen levels. The clot lysis rates, plasminogen, and plasma plasmin inhibitor levels returned to one-half their preinfusion level within 4–8 hr. The fibrinogen level returned to one-half its preinfusion level after 12 hr. Submitted on September 10, 1962

Interaction Between Low Molecular Weight Dextran Sulphate and Bentonite adsorbed Plasma

1979 ◽  
Author(s):  
T. Matsuda ◽  
M. Ogawara ◽  
T. Seki ◽  
R. Miura ◽  
M. Yokouchi ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Antithrombin Iii ◽  
Low Molecular Weight ◽  
Thrombin Time ◽  
Dextran Sulphate ◽  
Bothrops Atrox ◽  
Plasmin Inhibitor ◽  
Mean Molecular Weight

Effects of low molecular weight dextran sulphate (D.S.; mean molecular weight = 6,0006,500; S content = 17-19%) and/or bentonite (Wako Pure Chemical Industries, Osaka, Japan) adsorbed plasma (supernatant from plasma mixed with 300 mg/ml of bentonite for 10 minutes at 37°C), which contains no fibrinogen nor antithrombin III but α2 macro-globulin, α1-antitrypsin and α2-plasmin inhibitor, on thrombin time or batroxobin (purified fraction of venom of Bothrops atrox) time of fibrinogen solution were investigated. Addition of D.S. or bentonite adsorbed plasma to fibrinogen solution resulted in prolongations of thrombin time and batroxobin time. These results indicate that both D.S. and bentonite adsorbed plasma inhibit conversion of fibrinogen to fibrin by thrombin or batroxobin. However, when appropriately diluted bentonite adsorbed plasma and D.S. were added to fibrinogen solution simultaneously, thrombin times and batroxobin times were shorter compared to those when either bentonite adsorbed plasma or D.S. but not both was added. From these results, it is concluded that D.S. and bentonite adsorbed plasma interact to inhibit each other.

Changes in Fibrinogen After rt-PA Administration

1996 ◽  
Vol 2 (1) ◽  
pp. 43-50
Author(s):  
Justine Meehan Carr ◽  
Edwin G. Bovill ◽  
Russell P. Tracy ◽  
Martin Mankowski ◽  
Kenneth G. Mann ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Recombinant Tissue Plasminogen Activator ◽  
Low Molecular Weight ◽  
Tissue Plasminogen ◽  
Group A ◽  
Group B ◽  
Plasmin Inhibitor ◽  
D Dimer ◽  
Qualitative Changes ◽  
Time Point

Among patients participating in the TIMI-II protocol, there was a variability in the fibrinolytic re sponse to recombinant tissue plasminogen activator (rt- PA). A cohort of 20 TIMI-II patients was selected for detailed study because their responses to rt-PA varied widely in the degree of fibrin(ogen)olysis. Patient plasmas were analyzed by immunoblotting for changes in fibrino gen and plasminogen. Measurements of fibrinogen, fibrin ogen degradation product (FDP), D-dimer, Bβ 1-42, plas minogen, and t-PA were also correlated. Three patterns of response to rt-PA were identified: Group A ( n = 4) had fibrinogenolysis without fibrinolysis; Group B ( n = 11) had fibrinolysis and mild fibrinogenolysis; and Group C ( n = 5) had fibrinolysis with intense fibrinogenolysis. Group C patients also demonstrated qualitative changes in high- molecular-weight (HMW) and low-molecular-weight (LMW) fibrinogens, whereas Group A and B patients demonstrated only mild alterations in fibrinogen compo sition. Plasmin-inhibitor complexes were identified in all three groups. All patients had both plasmin-α2-anti plasmin and plasmin-α2-macroglobulin complexes at the 50-min time point. The concentration of pretreatment plasminogen correlated with the degree of fibrinogenoly sis.

Inhibition of the cathepsin B like proteinase by a low molecular weight cysteine-proteinase inhibitor from ascitic fluid and plasma α2 macroglobulin

1986 ◽  
Vol 64 (12) ◽  
pp. 1218-1225 ◽  
Author(s):  
Maurice Pagano ◽  
Daniel Keppler ◽  
Veronique Fumeron-Dalet ◽  
Robert Engler
Keyword(s):  
Molecular Weight ◽  
Ascitic Fluid ◽  
Cathepsin B ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Low Molecular Weight ◽  
Cysteine Proteinase Inhibitor ◽  
Cysteine Proteinase Inhibitors ◽  
Α2 Macroglobulin ◽  
Endopeptidase Activity

The cathepsin B like proteinase present in ascitic fluid of a patient with neoplasia has been purified and characterized after pepsin activation. From this fluid we have prepared the low molecular weight (LMW) cysteine-proteinase inhibitors. Three major inhibitor forms were found with isoelectric points of 7.4, 5.4, and 5.1, respectively. The interaction of the enzyme with the former inhibitor was studied because this inhibitor was the most abundant. The Ki value was found to be 4.3 × 10−8 M. Two molecules of this proteinase were bound by one molecule of plasma α2 macroglobulin (α2M). The LMW inhibitor was able to bind to the enzyme entrapped in α2M and reduced its endopeptidase activity on benzyloxycarbonyl-L-phenylalanyl-L-arginine-4-methyl-7-coumarylamide. These results may have a physiological significance in the regulation of the enzyme which, among other extracellular hydrolases, probably plays an important role in tumor invasion.

scholarly journals Crystal structure of a low molecular weight activator Blm-pep with yeast 20S proteasome – insights into the enzyme activation mechanism

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Julia Witkowska ◽  
Małgorzata Giżyńska ◽  
Przemysław Grudnik ◽  
Przemysław Golik ◽  
Przemysław Karpowicz ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Molecular Weight ◽  
Enzyme Activation ◽  
Activation Mechanism ◽  
Low Molecular Weight ◽  
20S Proteasome
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