• Clinical and Experimental Reproductive Medicine
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• v.20 no.1
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• pp.19-29
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• 1993

Plasminogen activator (PA)-plasmin system in follicular fluid is involved in the process leading to follicular rupture at ovulation. It is well known that PA is closely associated with cellular differentiation and tissue remodeling on evidences from the study of normal and malignant tissues. This study was designed to ascertain a potential role of PA in the ovarian folliculogenesis. Immature Sprague-Dawley rats were injected with pregnant mare serum gonadotropin, followed by injection of serine protease inhibitor (SPI; mixture of 1 mol/L benzamidine and 1 mol/L amino-caproic acid) into the unilateral ovarian bursa. In the control study, mechanical effect of bursal injection and contralateral ovarian effect SPI were ruled out. Total antral follicular areas relative to total ovarian cross-sectional areas was siginificantly lower in SPI-injected ovary than in saline-injected ovary. SPI injection decreased the relative antral follicular area by 33 % respectively. Electron microscopic finding of granulosa cell in the atretic follicle showed the presence of pyknotic nucleus, blurring of neucleolemma, degeneration of mitochondria and dilation of endoplasmic reticulum. After induction of ovulation with hCG, the number of oocytes released was significantly decreased in SPI-injected oviduct than in saline-injected oviduct. From above results, author discussed that PA may play a role not only in ovulation but also in some processes of folliculogenesis.

• The Korean Journal of Zoology
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• v.33 no.1
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• pp.119-125
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• 1990

A Potent inhibitor of trypsin and other various serine proteases including chymotrypsin, elastase, kallikrein, plasmin and subtilisin, has been purified to homogeneity from chick skeletal muscle by convendonal chromatographic procedures. The Inhibitor has an apparent molecular weight of 66, 000 dalton as determined by gel filtration. When the purified inhibitor was electrophoresed in the presence of sodium dodecyl sulfate, there appeared rwo protein bands having molecular weights of 66, 000 and 64, 000 dalton. The 64, 000 dalton protein seems to be the product of 66, 000 dalton protein by a lin'ited proteolysis during the purification procedure or in viuo. Thus, it seems to consist of a single polypeptide. The inhibitor appeared to be glycoprotein and have an isoelectric point of 7.4. It contains relatively large amount (8.33 mole%) of cysteine residues.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.107-114
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• 1985

In order to use for recipient strains of RP4:Mu cts, 5 strainsof Rhizobium were selected among 32 strains, which were isolated and identified in this study. Hybrid plasmin RP4::Mu cts, which, is temperature sensitive and confers resistance to ampicillin, kanamycin and tetracycline was transfered by conjugation from E. coli to other atrains of C. coli and the symbiotic nitrogen fixer, Rhizobium leguminosarum. Transfer frequencies of RP4::Mu cts plasmid from E. coli to Rhizobium were about $10^{-8}-10^{-7}$ in LB agar and YMA media. The transconjugants were confirmed by demonstrating that the drug-resistant and temperature-sensitive clones isolated were drug-resistant and temperature-sensitive clones isolated were capable of releasing phage and forming plaques. The plaque-forming units of transconjugants were about $10^2\;to\;10^3$. Stability test of RP4::Mucts in Rhizobium represented that most of the transconjugants had drug resistance and produce phage Mu cts.

• KSBB Journal
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• v.5 no.2
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• pp.183-189
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• 1990

An efficient method has been developed for the purification of urokinase from 1, 000 liter batches of human urine. The procedure involved precipitation of urokinase with 2mM zinc chloride, resuspension of the precipitate with 0.1M EDTA/0.5M Glycine solution, and CM-Toyopearl and benzamidine-Sepharose column chromatography. The purified urokinase was fully active and possessed a specific activity of 1.07$\times$105IU/mg. The recoveries ranged from 42 to 65% in several preparations(mean value was 51%). And the urokinase purified by this process consisted of about 13% of single chain urokinase (pro-urokinase) as evaluated by SDS-polyacrylamide gel electrophoresis in reducing condition and by S-2444 amldolytic activity under plasmin treatment.

• BMB Reports
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• v.47 no.12
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• pp.691-696
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• 2014

${\alpha}$-Enolase is a glycolytic enzyme and a surface receptor for plasminogen. ${\alpha}$-Enolase-bound plasminogen promotes tumor cell invasion and cancer metastasis by activating plasmin and consequently degrading the extracellular matrix degradation. Therefore, ${\alpha}$-enolase and plasminogen are novel targets for cancer therapy. We found that the amino acid sequence of a peptide purified from enzymatic hydrolysates of seahorse has striking similarities to that of ${\alpha}$-enolase. In this study, we report that this peptide competes with cellular ${\alpha}$-enolase for plasminogen binding and suppresses urokinase plasminogen activator (uPA)-mediated activation of plasminogen, which results in decreased invasive migration of HT1080 fibrosarcoma cells. In addition, the peptide treatment decreased the expression levels of uPA compared to that of untreated controls. These results provide new insight into the mechanism by which the seahorse-derived peptide suppresses invasive properties of human cancer cells. Our findings suggest that this peptide could emerge as a potential therapeutic agent for cancer.

• Bulletin of the Korean Chemical Society
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• v.30 no.1
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• pp.77-82
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• 2009

Thrombin activatable fibrinolysis inhibitor (TAFI) also known as plasma procarboxypeptidase B or U is a 60 kD glycoprotein, which is the major modulator of fibrinolysis in plasma. TAFI is a proenzyme, which is activated by proteolytic cleavage to an active carboxypeptidase B-like enzyme (TAFIa, 35.8 kD) by thrombin/thrombomodulin and plasmin. Modulation of fibrinolysis occurs when TAFIa enzymatically removes C-terminal lysine residues of partially degraded fibrin, thereby inhibiting the stimulation of tissue plasminogen activator (t-PA) modulated plasminogen activation. TAFIa undergoes a rapid conformational change at $37{^{\circ}C}$ to an inactive isoform called TAFIai. Potato tuber carboxypetidase inhibitor (PTCI) was shown to specifically bind to TAFIa as well as TAFIai. In this study, a novel immunoassay TAFIa/ai ELISA was used for quantitation of the two TAFI activation isoforms TAFIa and TAFIai. The ELISA utilizes PTCI as the capture agent and a double antibody sandwich technique for the detection. Low levels of TAFIa/ai antigen levels were detected in normal plasma and elevated levels were found in hemophilia A plasmas. TAFIa/ai antigen represents a novel marker to monitor fibrinolysis and TAFIa/ai ELISA may be a valuable assay for studying the role of TAFI in normal hemostasis and in pathological conditions.

• Archives of Pharmacal Research
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• v.21 no.4
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• pp.374-377
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• 1998

The freeze-dried powder of Lumbricus rubellus earthworm was administered orally to rats and its fibrinolytic and antithrombotic effects were investigated. The fibrinolytic activity of plasma was determined by measuring the plasmin activity of the euglobulin fraction and was increased to two-folds of the control at a dose of 0.5 g/kg/day and five times with 1 g/kg/day after 4-day administration. The antithrombotic effect was studied in an arterio-venous shunt model of rats. The thrombus weight decreased significantly from 43.2 mg to 32.4 mg at a dose of 0.5 g/kg/day after 8-day treatment. The level of fibrinogen/fibrin degradation product (FDP) in serum was elevated in a dose-dependent manner during the treatment period. On the 8th day after administration, the FDP value was increased to 7.7 $\mu\textrm{g}$g/ml compared with the control value of 3.3 $\mu\textrm{g}$g/ml. These results support that earthworm powder is valuable for the prevention and/or treatment of thrombotic conditions.

• Microbiology and Biotechnology Letters
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• v.51 no.2
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• pp.167-173
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• 2023

Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.12
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• pp.1708-1714
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• 2005

Keratinocyte growth factor (KGF) functions in epithelial growth and differentiation in many tissues and organs. KGF is expressed in the uterine endometrial epithelial cells during the estrous cycle and pregnancy in pigs, and receptors for KGF (KGFR) are expressed by conceptus trophectoderm and endometrial epithelia. KGF has been shown to stimulate the proliferation and differentiation of conceptus trophectoderm. However, the role of KGF on the endometrial epithelial cells has not been determined. Therefore, this study determined the effect of KGF on proliferation and differentiation of endometrial epithelial cells in vitro and in vivo using an immortalized porcine luminal epithelial (pLE) cell line and KGF infusion into the uterine lumen of pigs between Days 9 and 12 of estrous cycle. Results showed that KGF did not stimulate proliferation of uterine endometrial epithelial cells in vitro and in vivo determined by the $^3$H]thymidine incorporation assay and the proliferating cell nuclear antigen staining, respectively. Effects of KGF on expression of several markers for epithelial cell differentiation, including integrin receptor subunits $\alpha$4, $\alpha$5 and $\beta$1, plasmin/trypsin inhibitor, uteroferrin and retinol-binding protein were determined by RT-PCR, Northern and slot blot analyses, and immunohistochemisty, and KGF did not affect epithelial cell differentiation in vitro and in vivo. These results show that KGF does not induce epithelial cell proliferation and differentiation, suggesting that KGF produced by endometrial epithelial cells acts on conceptus trophectoderm in a paracrine manner rather than on endometrial epithelial cells in an autocrine manner.

• KSBB Journal
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• v.23 no.3
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• pp.251-256
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• 2008

The aim of this work was to investigate the fibrinolytic activity and characterization of Bacillus licheniformis HK-12, which produces the fibrinolytic enzyme excreted from naturally fermented Chungkook-Jang. Initially, the physiological and biochemical characteristics of strain HK-12 was examined. Both physiological analysis using BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were performed to identify the strain, and the strain could be assigned to Bacillus licheniformis, designated as B. lichenformis HK-12, and registered in GenBank as [EU288193]. Phylogenetic analysis of B. licheniformis HK-12 was plotted based on 16S rRNA sequence comparisons. During the incubation period of B. licheniformis HK-12, the changes of bacterial growth, fibrinolytic activity, and pH were monitored. As the results, after 36 hours of incubation, the maximum fibinolytic activity was about 2.25 times than that of plasmin used as standard. Optimal conditions on the growth of B. licheniformis HK-12 associated with the fibrinolytic activity was initial pH 7.0 and 40$^{\circ}C$, respectively.