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Die Medizinische Welt Oct 1961
Topics: Fibrinolysin; Thromboembolism
PubMed: 14494137
DOI: No ID Found -
Proceedings of the National Academy of... May 1979Plasminogen was found to be present in bovine milk by crossreactivity between rabbit antiserum to plasminogen and casein prepared from milk by acid precipitation. This...
Plasminogen was found to be present in bovine milk by crossreactivity between rabbit antiserum to plasminogen and casein prepared from milk by acid precipitation. This result was further supported by recovery of intact 125I-labeled plasminogen from rabbit milk after its intravenous injection. Freshly isolated whole bovine casein was observed to undergo slow autoproteolysis at 37 degrees C. Polyacrylamide gel electrophoresis revealed gradual disappearance of major caseins accompanied by appearance and increase in intensity of numerous electrophoretic bands. This autoproteolysis was inhibited by low concentrations of epsilon-aminocaproic acid (0.1 mM) and diisopropyl fluorophosphate (1 mM); catalytic amounts of urokinase accelerated the process. Autoproteolysis of isolated bovine beta-casein was shown by both urea and sodium dodecyl sulfate gel electrophoresis to result in formation of gamma 1- and gamma 2-caseins. Similar electrophoretic bands were formed when beta-casein was degraded by plasmin prepared from bovine blood serum. These results support the hypothesis that bovine plasmin occurs in milk and is identical to alkaline milk protease.
Topics: Animals; Caseins; Cattle; Female; Fibrinolysin; Hydrolysis; Milk; Molecular Weight; Plasminogen; Protease Inhibitors
PubMed: 156365
DOI: 10.1073/pnas.76.5.2244 -
ACS Applied Materials & Interfaces Jan 2023The fibrillization and deposition of the human islet amyloid polypeptide (hIAPP) are the pathological hallmark of type 2 diabetes mellitus (T2DM), and these insoluble...
The fibrillization and deposition of the human islet amyloid polypeptide (hIAPP) are the pathological hallmark of type 2 diabetes mellitus (T2DM), and these insoluble fibrotic depositions of hIAPP are considered to strongly affect insulin secretion by inducing toxicity toward pancreatic islet β-cells. The current strategy of preventing amyloid aggregation by nanoparticle-assisted inhibitors can only disassemble fibrotic amyloids into more toxic oligomers and/or protofibrils. Herein, for the first time, we propose a type of cysteine-derived chiral carbon quantum dot (CQD) that targets plasmin, a core natural fibrinolytic protease in humans. These CQDs can serve as fibrinolytic activity regulators for plasmin to cleave hIAPP into nontoxic polypeptides or into even smaller amino acid fragments, thus alleviating hIAPP's fibrotic amyloid-induced cytotoxicity. Our experiments indicate that chiral CQDs have opposing effects on plasmin activity. The l-CQDs promote the cleavage of hIAPP by enhancing plasmin activity at a promotion ratio of 23.2%, thus protecting β-cells from amyloid-induced toxicity. In contrast, the resultant d-CQDs significantly inhibit proteolysis, decreasing plasmin activity by 31.5% under the same reaction conditions. Second harmonic generation (SHG) microscopic imaging is initially used to dynamically characterize hIAPP before and after proteolysis. The l-CQD promotion of plasmin activity thus provides a promising avenue for the hIAPP-targeted treatment of T2DM to treat low fibrinolytic activity, while the d-CQDs, as inhibitors of plasmin activity, may improve patient survival for hyperfibrinolytic conditions, such as those existing during surgeries and traumas.
Topics: Humans; Amyloid; Carbon; Cysteine; Diabetes Mellitus, Type 2; Fibrinolysin; Islet Amyloid Polypeptide; Quantum Dots
PubMed: 36596222
DOI: 10.1021/acsami.2c17975 -
Journal of Enzyme Inhibition and... Aug 2012Plasmin plays important roles in various physiological systems. The identification of inhibitors controlling its regulation represents a promising drug-discovery...
Plasmin plays important roles in various physiological systems. The identification of inhibitors controlling its regulation represents a promising drug-discovery challenge. To develop selective inhibitors of plasmin, structural information of the binding modes is crucial. Here, a computational docking study was conducted to provide structural insight into plasmin subsite interactions with substrates/inhibitors. Predicted binding modes of two peptide-substrates (D/L-Ile-Phe-Lys), and potent and weak inhibitors (YO-2 and PKSI-527) suggested non-prime and prime subsite interactions relevant to recognition by plasmin. Predicted binding modes also correlated well with the experimental structure-activity relationships for plasmin substrates/inhibitors, namely the differences of K(M) values between the D- and L-peptide-substrates and inhibitory potencies of YO-2 and PKSI-527. In particular, interaction observed at a hydrophobic pocket near S2 and at a tunnel-shaped hydrophobic S1' was strongly suggested to be significantly involved in tight binding of inhibitors to plasmin. Our present findings may aid in the design of potent and selective plasmin inhibitors.
Topics: Computational Biology; Dipeptides; Dose-Response Relationship, Drug; Fibrinolysin; Models, Molecular; Molecular Structure; Oligopeptides; Phenylalanine; Serine Proteinase Inhibitors; Structure-Activity Relationship; Substrate Specificity; Tranexamic Acid
PubMed: 21992704
DOI: 10.3109/14756366.2011.603129 -
Journal of Dairy Science Nov 1993Bovine plasmin (EC 3.4.21.7) activity was measured on H-D-valyl-L-leucyl-L-lysyl-4-nitroanilide and acid casein in the presence of native and heat-denatured...
Bovine plasmin (EC 3.4.21.7) activity was measured on H-D-valyl-L-leucyl-L-lysyl-4-nitroanilide and acid casein in the presence of native and heat-denatured beta-lactoglobulin (denatured at 100 degrees C for 15 min before being mixed with plasmin solutions). Native or denatured beta-lactoglobulin was then heated with plasmin at 60 degrees C for 15 min. Enzyme activity again was estimated after this mild heat treatment. Native and denatured beta-lactoglobulin inhibited the action of plasmin on H-D-valyl-L-leucyl-L-lysyl-4-nitroanilide and casein. The mild heat treatment (60 degrees C for 15 min) caused stronger inhibition of the activity of plasmin against casein and the synthetic substrate. For H-D-valyl-L-leucyl-L-lysyl-4-nitroanilide, inhibition was competitive in unheated mixtures, but heating beta-lactoglobulin with plasmin changed inhibition type to mixed. This change suggests a heat-dependent interaction between plasmin and beta-lactoglobulin. Native beta-lactoglobulin was more inhibitory of plasmin's action against casein than was denatured beta-lactoglobulin. The converse was observed when plasmin activity was measured with the synthetic substrate.
Topics: Animals; Caseins; Cattle; Fibrinolysin; Hot Temperature; Kinetics; Lactoglobulins; Micelles; Oligopeptides; Protein Denaturation
PubMed: 8270680
DOI: 10.3168/jds.S0022-0302(93)77673-0 -
The Journal of Clinical Investigation Mar 1970Plasmin incubated with partially purified C[unk] inactivator produced a decrease in inhibitory activity which was related to the time of incubation and to the...
Plasmin incubated with partially purified C[unk] inactivator produced a decrease in inhibitory activity which was related to the time of incubation and to the concentration of plasmin. This effect of plasmin was not influenced by the purity of the inhibitor preparations. Soybean trypsin inhibitor and tosyl arginine methyl ester (TAMe), substances which block the active enzymic center of plasmin, prevented the plasmin-induced inactivation. Double diffusion analysis of the functionally deficient, plasmin-treated C[unk] inactivator using a specific antibody, showed a reaction of identity with the untreated inhibitor. Agarose and acrylamide gel immunoelectrophoresis of a plasmin, inhibitor mixture showed the appearance of an additional precipitin band with immunologic reactivity similar to that of the untreated inhibitor. These results demonstrate that plasmin alters both the functional and immunoelectrophoretic properties of C[unk] inactivator, and that the active proteolytic site of plasmin is necessary for this interaction. Since C[unk] inactivator has been shown to inhibit several different proteolytic enzymes including C[unk], kallikrein, PF/Dil, and plasmin, this investigation provides a theoretical relationship between the fibrinolytic, kallikrein, and complement systems which may have pathophysiologic relevance to various human disease states.
Topics: Arginine; Chromatography, Thin Layer; Complement System Proteins; Esters; Fibrinolysin; Immunodiffusion; Immunoelectrophoresis; Kallikreins; Sulfonic Acids; Trypsin Inhibitors
PubMed: 4244455
DOI: 10.1172/JCI106267 -
The Journal of Biological Chemistry Jul 1991The purpose of this investigation was to characterize the reaction of alpha 2-antiplasmin (alpha 2AP) and alpha 2-macroglobulin (alpha 2M) with human plasmin bound to...
The purpose of this investigation was to characterize the reaction of alpha 2-antiplasmin (alpha 2AP) and alpha 2-macroglobulin (alpha 2M) with human plasmin bound to rat C6 glioma cells and human umbilical vein endothelial cells (HUVECs). Binding of plasmin (0.1 microM) to C6 cells at 4 degrees C did not cause cell detachment, decrease viability or change cell morphology. The KD and Bmax for the binding of diisopropyl phosphoryl plasmin (DIP-plasmin) to C6 cells were 0.9 microM and 2.6 x 10(6) sites/cell. The dissociation rate constants (koff) for 125I-plasmin were 9.7 x 10(-4) and 4.0 x 10(-4) s-1 at 4 degrees C in the presence and absence of 0.3 microM DIP-plasmin, respectively. Similar constants were determined for 125I-plasminogen and 125I-DIP-plasmin. Neither alpha 2AP nor alpha 2M affected the dissociation of DIP-plasmin. C6 cell-associated 125I-plasmin reacted slowly with alpha 2AP; however, the inhibition rate constants exceeded the koff. alpha 2AP-plasmin complex formed after the plasmin dissociated into solution (reaction pathway 1) and by direct reaction of alpha 2AP with cell-associated enzyme (reaction pathway 2). High concentrations of alpha 2AP favored pathway 2. C6 cell-associated plasmin was also protected from inhibition by alpha 2M. While the same pathways were probably involved in this reaction, alpha 2M was less effective than alpha 2AP as an inhibitor of nondissociated plasmin (pathway 2). When C6 cell-bound plasmin reacted with alpha 2AP, alpha 2AP-plasmin complex was recovered primarily in the medium, suggesting dissociation of complexes formed on the cell surface. Plasmin-receptor dissociation and inhibition experiments were performed at 22 degrees and 37 degrees C, confirming the conclusions of the 4 degrees C studies. Comparable results were also obtained using HUVEC cultures. These studies demonstrate that cell-associated plasmin is protected from inhibition by alpha 2M as well as alpha 2AP. At least two reaction pathways may be demonstrated for the inhibition of plasmin that is initially receptor-bound; however, neither pathway is highly effective, accounting for the "plasmin-protective" activity of the cell surface.
Topics: Animals; Aprotinin; Electrophoresis, Polyacrylamide Gel; Fibrinolysin; Humans; Plasminogen; Rats; Receptors, Cell Surface; Receptors, Peptide; Substrate Specificity; Temperature; Tumor Cells, Cultured; alpha-Macroglobulins
PubMed: 1712017
DOI: No ID Found -
Investigative Ophthalmology & Visual... Sep 2004To evaluate the safety and efficacy of dispase and plasmin when inducing posterior vitreous detachment (PVD) by intravitreous injection in rabbit eyes. (Comparative Study)
Comparative Study
PURPOSE
To evaluate the safety and efficacy of dispase and plasmin when inducing posterior vitreous detachment (PVD) by intravitreous injection in rabbit eyes.
METHODS
Forty-eight young pigmented rabbits were randomized into six groups. Groups 1 and 5 received 0.025 U dispase in test eyes; group 2, 0.1 U dispase; groups 3 and 6, 1 U plasmin; and group 4, 4 U plasmin. All groups received PBS in control eyes. Groups 5 and 6 were euthanatized 15 minutes after surgery for ocular histologic examination. The remaining groups (groups 1-4) received indirect ophthalmoscope and biomicroscopy 15 and 30 minutes; 1, 2, and 8 hours; and 1, 3, and 7 days after surgery. Ultrasonography and electroretinogram were performed 1 hour and 1 and 7 days after surgery. The eyes then were examined by scanning and transmission electron microscopy.
RESULTS
Partial or complete PVDs were observed in the eyes that received dispase and plasmin, confirmed by the results of scanning electron microscopy. Light microscopy showed inflammation in both dispase- and plasmin-treated eyes of groups 5 and 6. However, whereas in plasmin-treated eyes the ERG and cell ultrastructure showed no significant changes, in dispase-treated eyes, the amplitudes of ERG showed a significant reduction from baseline and ultrastructural damage to the retina was detected by transmission electron microscopy. Cell damage, preretinal hemorrhage, and cataract were also observed in these eyes. No changes were observed in the control eyes.
CONCLUSIONS
Intravitreal injection of dispase at 0.025 U or more can induce PVD, but it is not safe. Plasmin (1-4 U) is safer, except for the potential risk of inducing intraocular inflammation.
Topics: Animals; Cataract; Dose-Response Relationship, Drug; Electroretinography; Endopeptidases; Endophthalmitis; Fibrinolysin; Fundus Oculi; Injections; Microscopy, Electron; Microscopy, Electron, Scanning; Rabbits; Random Allocation; Retinal Hemorrhage; Ultrasonography; Vitreous Body
PubMed: 15326153
DOI: 10.1167/iovs.04-0026 -
Theriogenology Aug 2004Proteolytic enzymes appear to have an essential role in multiple phases of mammalian fertilization. Several observations suggest that the plasminogen activator/plasmin...
Proteolytic enzymes appear to have an essential role in multiple phases of mammalian fertilization. Several observations suggest that the plasminogen activator/plasmin system might also play a role in mammalian fertilization. Movement characteristics of bovine sperm incubated with different concentrations of plasmin were investigated using a computer-assisted automated semen analysis system. Sperm were incubated up to 4h in a modified Tyrode's medium (control) and 0.1, 1, 10 and 100 mU/ml of plasmin. The percentage motile sperm was significantly higher at 0 h for sperm incubated in 1, 10, and 100 mU of plasmin. Relative to sperm incubated in control medium, lateral head displacement (ALH), curvilinear velocity, beat cross frequency, path velocity and straight line velocity (VSL) of sperm treated with 100 mU of plasmin for 0 h were increased. After 2h of incubation, sperm treated with 100 mU of plasmin showed an increase in ALH, but a decrease in VSL, straightness and linearity. The effect of plasmin on most motility parameters appears to be direct since all these parameters were affected at 0 h of incubation. Our results support the notion of hyperactivation of bovine spermatozoa following incubation with different concentrations of plasmin. The present work provides additional information to further characterize motility movement of bovine sperm associated with final preparation for fertilization.
Topics: Animals; Cattle; Fertilization; Fibrinolysin; Male; Sperm Motility; Spermatozoa
PubMed: 15226011
DOI: 10.1016/j.theriogenology.2003.11.016 -
Bioorganic & Medicinal Chemistry Feb 2016Based on the structure of YO-2 [N-(trans-4-aminomethylcyclohexanecarbonyl)-l-Tyr(O-picolyl)-NH-octyl], active site-directed plasmin (Plm) inhibitors were explored. The...
Based on the structure of YO-2 [N-(trans-4-aminomethylcyclohexanecarbonyl)-l-Tyr(O-picolyl)-NH-octyl], active site-directed plasmin (Plm) inhibitors were explored. The picolyl moiety in the Tyr(O-picolyl) residue (namely, the P2 residue) was replaced with smaller or larger groups, such as hydrogen, tert-butyl, benzyl, (2-naphthyl)methyl, and (quinolin-2-yl)methyl. Those efforts produced compound 17 {N-(trans-4-aminomethylcyclohexanecarbonyl)-l-Tyr[O-(quinolin-2-yl)methyl]-NH-octyl} [IC50=0.22 and 77μM for Plm and urokinase (UK), respectively], which showed not only 2.4-fold greater Plm inhibition than YO-2, but also an improvement in selectivity (Plm/UK) by 35-fold. The docking experiments of the Plm-17 complexes disclosed that the amino group of the tranexamyl moiety interacted with the side-chain of Asp753 which formed S1 site.
Topics: Antifibrinolytic Agents; Catalytic Domain; Dose-Response Relationship, Drug; Fibrinolysin; Humans; Molecular Docking Simulation; Molecular Structure; Structure-Activity Relationship; Tyrosine; Urokinase-Type Plasminogen Activator
PubMed: 26732532
DOI: 10.1016/j.bmc.2015.12.009