-
Pulmonary Pharmacology & Therapeutics Oct 2021
Topics: Benzamidines; Fibrinolysin; Guanidines; Humans; COVID-19 Drug Treatment
PubMed: 34271164
DOI: 10.1016/j.pupt.2021.102055 -
Arteriosclerosis, Thrombosis, and... May 2017Previous studies have demonstrated a role for plasmin in regulating plasma von Willebrand factor (VWF) multimer composition. Moreover, emerging data have shown that...
OBJECTIVE
Previous studies have demonstrated a role for plasmin in regulating plasma von Willebrand factor (VWF) multimer composition. Moreover, emerging data have shown that plasmin-induced cleavage of VWF is of particular importance in specific pathological states. Interestingly, plasmin has been successfully used as an alternative to ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 1 motif) in a mouse model of thrombotic thrombocytopenic purpura. Consequently, elucidating the molecular mechanisms through which plasmin binds and cleaves VWF is not only of basic scientific interest but also of direct clinical importance. Our aim was to investigate factors that modulate the susceptibility of human VWF to proteolysis by plasmin.
APPROACH AND RESULTS
We have adapted the VWF vortex proteolysis assay to allow for time-dependent shear exposure studies. We show that globular VWF is resistant to plasmin cleavage under static conditions, but is readily cleaved by plasmin under shear. Although both plasmin and ADAMTS13 cleave VWF in a shear-dependent manner, plasmin does not cleave at the Tyr1605-Met1606 ADAMTS13 proteolytic site in the A2 domain. Rather under shear stress conditions, or in the presence of denaturants, such as urea or ristocetin, plasmin cleaves the K1491-R1492 peptide bond within the VWF A1-A2 linker region. Finally, we demonstrate that VWF susceptibility to plasmin proteolysis at K1491-R1492 is modulated by local N-linked glycan expression within A1A2A3, and specifically inhibited by heparin binding to the A1 domain.
CONCLUSIONS
Improved understanding of the plasmin-VWF interaction offers exciting opportunities to develop novel adjunctive therapies for the treatment of refractory thrombotic thrombocytopenic purpura.
Topics: Binding Sites; Fibrinolysin; Heparin; Humans; Polysaccharides; Protein Binding; Protein Interaction Domains and Motifs; Proteolysis; Stress, Mechanical; Structure-Activity Relationship; Time Factors; von Willebrand Factor
PubMed: 28279966
DOI: 10.1161/ATVBAHA.116.308524 -
Acta Ophthalmologica. Supplement 1992The involvement and role of the plasminogen activator-plasmin system in normal and pathological wound healing is reviewed. The methods currently available for... (Review)
Review
The involvement and role of the plasminogen activator-plasmin system in normal and pathological wound healing is reviewed. The methods currently available for demonstrating plasmin activity are briefly described. The article also reviews some other serine proteases potentially involved in pathological wound healing processes. The current ophthalmological therapeutic measures used to regulate tissue proteolysis are also described.
Topics: Cornea; Corneal Ulcer; Fibrinolysin; Humans; Plasminogen; Plasminogen Activators; Serine Endopeptidases; Tears; Wound Healing
PubMed: 1322011
DOI: 10.1111/j.1755-3768.1992.tb02168.x -
The Journal of Biological Chemistry Sep 2004Plasmin is a major extracellular protease that elicits intracellular signals to mediate platelet aggregation, chemotaxis of peripheral blood monocytes, and release of...
Plasmin is a major extracellular protease that elicits intracellular signals to mediate platelet aggregation, chemotaxis of peripheral blood monocytes, and release of arachidonate and leukotriene from several cell types in a G protein-dependent manner. Angiostatin, a fragment of plasmin(ogen), is a ligand and an antagonist for integrin alpha(9)beta(1). Here we report that plasmin specifically interacts with alpha(9)beta(1) and that plasmin induces of cells expressing migration recombinant alpha(9)beta(1) (alpha(9)-Chinese hamster ovary (CHO) cells). Migration was dependent on an interaction of the kringle domains of plasmin with alpha(9)beta(1) as well as the catalytic activity of plasmin. Angiostatin, representing the kringle domains of plasmin, alone did not induce the migration of alpha(9)-CHO cells, but simultaneous activation of the G protein-coupled protease-activated receptor (PAR)-1 with an agonist peptide induced the migration on angiostatin, whereas PAR-2 or PAR-4 agonist peptides were without effect. Furthermore, a small chemical inhibitor of PAR-1 (RWJ 58259) and a palmitoylated PAR-1-blocking peptide inhibited plasmin-induced migration of alpha(9)-CHO cells. These results suggest that plasmin induces migration by kringle-mediated binding to alpha(9)beta(1) and simultaneous proteolytic activation of PAR-1.
Topics: Animals; CHO Cells; Catalysis; Cell Movement; Cricetinae; Fibrinolysin; Humans; Integrins; Protein Binding; Receptor, PAR-1; Signal Transduction
PubMed: 15247268
DOI: 10.1074/jbc.M401372200 -
The Biochemical Journal Jul 1990The kinetics of inhibition of the amidolytic activity of plasmin on D-Val-L-Leu-L-Lys p-nitroanilide hydrochloride (S-2251) by fibrinogen and fibrin were determined....
The kinetics of inhibition of the amidolytic activity of plasmin on D-Val-L-Leu-L-Lys p-nitroanilide hydrochloride (S-2251) by fibrinogen and fibrin were determined. Reciprocal (1/v versus 1/[S]) plots of plasmin inhibition by 0.50 microM-fibrinogen showed a non-linear downward curve. The Hill coefficient (h) was 0.68, suggesting negative co-operativity. By contrast, fibrin produced a simple competitive inhibition of plasmin (Ki = 12 micrograms/ml). Addition of 0.1 mM-6-aminohexanoic acid shifted the non-linear curve obtained in the presence of fibrinogen to a straight line as for controls, indicating that 6-aminohexanoic acid abolishes the fibrinogen-induced inhibition. Transient exposure of the enzyme to pH 1.0 abrogates the ability of fibrinogen to inhibit plasmin activity. Acidification had no effect on the Vmax but increased the Km of plasmin. The present evidence for modulation of plasmin reveals a novel mechanism for control of fibrinolysis by fibrinogen, a component of the coagulation system and the precursor of the physiological substrate of plasmin.
Topics: Amino Acid Sequence; Aminocaproic Acid; Binding, Competitive; Chromogenic Compounds; Fibrin; Fibrinogen; Fibrinolysin; Hydrogen-Ion Concentration; Kinetics; Molecular Sequence Data; Oligopeptides
PubMed: 2143650
DOI: 10.1042/bj2690299 -
Nihon Rinsho. Japanese Journal of... Sep 1999
Review
Topics: Antifibrinolytic Agents; Fibrinolysin; Humans; alpha-2-Antiplasmin
PubMed: 10543213
DOI: No ID Found -
Acta Ophthalmologica Dec 1988Proteolytic activity was studied in subretinal fluid from 56 eyes with rhegmatogenous retinal detachment without vitreous or subretinal hemorrhage. Active plasmin...
Proteolytic activity was studied in subretinal fluid from 56 eyes with rhegmatogenous retinal detachment without vitreous or subretinal hemorrhage. Active plasmin (1.0-15.2 micrograms/ml) was found in 33 eyes and plasmin-inhibitor complexes in 3 eyes. Plasmin was detected more often in large detachments, but there was no clear correlation with the duration of the detachment or characteristics of the holes in the retina. It seems possible that plasmin in subretinal fluid may enhance release of cells from the pigment epithelium by degrading the extracellular matrix and contribute to the development of proliferative vitreoretinopathy.
Topics: Electrophoresis, Polyacrylamide Gel; Fibrinolysin; Peptide Hydrolases; Plasminogen; Retina; Retinal Detachment
PubMed: 2976571
DOI: 10.1111/j.1755-3768.1988.tb04055.x -
Biochemical and Biophysical Research... Jul 1991The rate of plasmin denaturation was in the order of Lys-plasmin greater than miniplasmin greater than microplasmin. Fibrinogen degradation products (FDP) dose...
The rate of plasmin denaturation was in the order of Lys-plasmin greater than miniplasmin greater than microplasmin. Fibrinogen degradation products (FDP) dose dependently increased the denaturation rate of Lys-plasmin and mini-plasmin with a maximal rate constant at the FDP/plasmin ratio of about 0.5. The denaturation rate constant of microplasmin was not affected. FDP increased the rate of plasmin denaturation was in parallel with its effect on the interaction among kringle domains. Without FDP only trace amounts of plasminogen dimer could be detected by cross-linking with bis-(sulfo-succinimidyl)-suberate followed by SDS gel electrophoresis. In the low concentration of FDP significant amounts of oligomers of Glu-, mini-plasminogens, kringle 1-3 and kringle 1-5 were observed. High concentration of FDP, however, decreased plasminogen oligomer.
Topics: Amino Acid Sequence; Cross-Linking Reagents; Fibrinolysin; Humans; Kinetics; Models, Biological; Molecular Weight; Plasminogen; Protein Conformation; Protein Denaturation; Succinimides
PubMed: 1829887
DOI: 10.1016/0006-291x(91)91822-t -
Biochemical and Biophysical Research... Feb 2015The potent fibrinolytic enzyme, plasmin has numerous clinical applications for recannulizing vessels obstructed by thrombus. Despite its diminutive size, 91 kDa, success... (Comparative Study)
Comparative Study
The potent fibrinolytic enzyme, plasmin has numerous clinical applications for recannulizing vessels obstructed by thrombus. Despite its diminutive size, 91 kDa, success in the recombinant expression of this serine protease has been limited. For this reason, a truncated non-glycosylated plasmin variant was developed capable of being expressed and purified from E. coli. This mutated plasmin, known as δ-plasmin, eliminates four of the five kringle domains present on native plasmin, retaining only kringle 1 fused directly to the unmodified catalytic domain of plasmin. This study demonstrates that δ-plasmin exhibits similar kinetic characteristics to full length plasmin despite its heavily mutated form; KM = 268.78 ± 19.12, 324.90 ± 8.43 μM and Kcat = 770.48 ± 41.73, 778.21 ± 1.51 1/min for plasmin and δ-plasmin, respectively. A comparative analysis was also carried out to investigate the inhibitory effects of a range of benzamidine based small molecule inhibitors: benzamidine, p-aminobenzamidine, 4-carboxybenzamidine, 4-aminomethyl benzamidine, and pentamidine. All of the small molecule inhibitors, with the exception of unmodified benzamidine, demonstrated comparable competitive inhibition constants (Ki) for both plasmin and δ-plasmin ranging from Ki < 4 μM for pentamidine to Ki > 1000 μM in the case of aminomethyl benzamidine. This result further supports that δ-plasmin retains much of the same functionality as native plasmin despite its greatly reduced size and complexity. This study serves the purpose of demonstrating the tunable inhibition of plasmin and δ-plasmin with potential applications for the improved clinical delivery of δ-plasmin to treat various thrombi.
Topics: Antifibrinolytic Agents; Benzamidines; Fibrinolysin; Humans; Kinetics; Models, Molecular; Mutant Proteins; Plasminogen; Protein Conformation; Protein Structure, Tertiary; Recombinant Proteins; Structure-Activity Relationship
PubMed: 25576865
DOI: 10.1016/j.bbrc.2014.12.117 -
Biochemical and Biophysical Research... Apr 2019Plasmin is a potent serin protease involved in a variety of biological functions, such as fibrinolysis and tissue remodeling. On performing an in vitro control assay to...
Plasmin is a potent serin protease involved in a variety of biological functions, such as fibrinolysis and tissue remodeling. On performing an in vitro control assay to measure the activity of endogenous plasmin in cell lysates, a stimulatory effect of non-ionic detergent NP-40 on plasmin activity was discovered. Another non-ionic detergent, TX-100, also enhanced plasmin activity, while ionic detergents sodium deoxycholate and sodiem dodecyl sulfate abolished plasmin enzyme activity. Kinetic analysis of plasmin activity in the presence of NP-40 and TX-100 demonstrated an increase in V; however, there was no change in K values, suggesting that these detergents stimulate plasmin activity in a non-competitive manner. Fibrin plate assay indicates that NP-40 and TX-100 functionally stimulate plasmin activity by showing a dose-dependent increase in fibrinolysis.
Topics: Deoxycholic Acid; Detergents; Fibrinolysin; Fibrinolysis; Humans; In Vitro Techniques; Kinetics; Octoxynol; Sodium Dodecyl Sulfate
PubMed: 30890336
DOI: 10.1016/j.bbrc.2019.03.052