• Molecules and Cells
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• v.24 no.2
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• pp.240-246
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• 2007

The ${\beta}2$ integrins on leukocytes play important roles in cell adhesion, migration and phagocytosis. One of the ${\beta}2$ integrins, ${\alpha}X{\beta}2$ (CD11c/CD18), is known to bind ligands such as fibrinogen, Thy-1 and iC3b, but its function is not well characterized. To understand its biological roles, we attempted to identify novel ligands. The functional moiety of ${\alpha}X{\beta}2$, the ${\alpha}X$ I-domain, was found to bind plasminogen, the zymogen of plasmin, with moderate affinity ($1.92{\times}10^{-6}M$) in the presence of $Mg^{2+}$ or $Mn^{2+}$. The ${\beta}D-{\alpha}5$ loop of the ${\alpha}X$ I-domain proved to be responsible for binding, and lysine residues ($Lys^{242}$, $Lys^{243}$) in the loop were the most important for recognizing plasminogen. An excess amount of the lysine analog, 6-aminohexanoic acid, inhibited ${\alpha}X$ I-domain binding to plasminogen, indicating that binding is lysine-dependent. The results of this study indicate that leukocytes regulate plasminogen activation, and consequently plasmin activities, through an interaction with ${\alpha}X{\beta}2$ integrin.

• The Korean Journal of Zoology
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• v.31 no.3
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• pp.185-190
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• 1988

To investigate whether malignant behavior of skin tumor correlates with changes in the level of proteolytic activities in skin tumors, rats were treated with 7,12-dimethylbenzanthracene followed by photbor ester. Tumors induced upon the treatrnents exhibited more than 20-fold increase in the activity of plasminogen activator and about 3-fold of plasmin-like activity. as compared to those in treated controls. Furthermore, the former activity was raised to about 6-fold even in the preneoplastic dssues of the skin tissues. On the other hand, the proteolytic activity against casein and insulin decreased to several-fold in the tumor tissues while antitrvpsin activity remained similar in both tumor and controls. Thus, the increase in the activities of plasmInogen activator and plasminlike enzyme appears to occur as a charaderistic to skin cancer and may involve in invasion and metsstasis of the tumor.

• Journal of Mushroom
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• v.13 no.3
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• pp.192-198
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• 2015

This study was performed in order to analyze the fibrinolytic, thrombin inhibitory, anti-oxidative, acetylcholinesterase inhibitory, and immuno-enhancing activities of the water extract and solvent fractions isolated from Grifola frondosa. Fibrinolytic activity was analyzed using the fibrin plate method, and thrombin inhibitory activity was assayed using the substrate H-D-Phe-piparg- pna. Anti-oxidative activity was estimated using the DPPH assay, and AChE inhibitory activity was measured using the spectrophotometric method. Immuno-enhancing activity was examined using the nitric oxide (NO) production in RAW 264.7 macrophage cells. Cell viability was determined using the MTS assay. Fibrinolytic activities were the highest in water extract (1.55 plasmin units/mL) followed by water fraction (0.85 plasmin units/mL). The thrombin inhibitory activities of the water and ethyl acetate fractions were determined to be 76.43% and 72.59%, respectively. The acetylcholinesterase inhibitory activities of chloroform and hexane fractions exhibited values of 95.14% and 94.74%, respectively. The butanol fraction showed the highest anti-oxidative activity at 94.47%. Anti-proliferating activity against Raw 264.7 cells showed no cytotoxicity. The production of NO in Raw 264.7 cells increased up to 2-fold by adding the water fraction compared to the untreated control. These results suggest that Grifola frondosa may serve as a useful functional food for the enhancement of immune function and the prevention and therapy of cardiovascular diseases.

• BMB Reports
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• v.34 no.2
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• pp.134-138
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• 2001

By adding sodium chloride (2.5%) into a Bacillus amyloliquefaciens DJ-4 culture broth, two serine-type fibrinolytic proteases with a molecular weight of 29 (subtilisin DJ-4) and 38-kDa were stimulated on the SDS-fibrin zymogram or inhibitor gels. B. amyloliquefaciens DJ-4 showed the highest proteolytic activity (5.52 plasmin NIH unit/ml) on the fibrin plate based on the molar ratio when cells were subjected to the 2.5% NaCl. Using a fibrin plate, the secreted protease from this strain in the presence of 5% NaCl showed that about 49% of the enzyme's activity remained after incubation at $60^{\circ}C$ for 30 min, but as the salt concentration was increased (10% NaCl) the activity nearly disappeared (0.14 plasmin NIH unit/ml). However, through a fibrin zymography assay, three fibrinolytic enzymes (38, 53 and 80-kDa) from the cells in the presence of 10% NaCl were detected. Also, two salt-activated serine-type fibrinolytic professes (29 and 38kDa) showed thermostability from 65 to $70^{\circ}C$ for 30 min. Furthermore, these professes also showed stability, pH 6-11. In particular, 29-kDa (subtilisin DJ-4) was very stable in the pH range of 4-11 at $4^{\circ}C$ for 48 h.

• Reproductive and Developmental Biology
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• v.38 no.2
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• pp.63-70
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• 2014

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. Two type of PAs are urokinase-type PA (uPA) and tissue-type PA (tPA). Plasminogen is present in most extracellular fluids. PAs play in various reproductive processes including implantation, ovulation and fertilization. In the spermatozoa, PAs and PAIs play a role in sperm motility and fertilization. PAs in the sertoli cell are stimulated spermatozoa maturation and sperm activation through the phospholipase A2. The oocyte maturation is the process for fertilization and implantation. PAs in cumulus-oocyte complexes (COCs) are related to oocyte maturation by protein kinase A and C. In the ovulatory process, PAs activity are changed and it are related to reducing the tensile strength of ovarian follicle wall. The uterine environment is important for reproduction and the uterus undergo tissue remodeling. In the uterus and oviduct of mammals, expression and activity of PAs are changed during estrous cycle. Thus, expression and activity of PAs are concerned to many reproductive functions. Therefore, PAs seem to important factor of regulator in reproductive events.

• Biomedical Science Letters
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• v.12 no.4
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• pp.393-398
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• 2006

These studies were carried out to characterize the physiological functionalities of Codonopsis lanceolata, Cornus officinalis, and their mixtures. We investigated the antioxidative, fibrinolytic, and ${\alpha}$-glucosidase inhibitory activities of them. The antioxidative activities of Codonopsis lanceoiata and Cornus officinalis were 87% and 90%, respectively. Addition of salt to Codonopsis lanceolata and Cornus officinalis did not affect its antioxidative activities. In spite of fourfold addition of Codonopsis lanceolata to Cornus officinalis, the antioxidative activity was conserved at 90%. The fibrinolytic activities of Codonopsis lanceolata and Cornus officinalis were 0.78 plasmin unit/ml and 1.74 plasmin unit/ml, respectively. Addition of salt decreased the fibrinolytic activities of both Codonopsis lanceolata and Cornus officinalis. A mixture (3:1) of Codonopsis lanceolata and Cornus officinalis exhibited a 21% increase in activity. The ${\alpha}$-glucosidase inhibitory activities of Codonopsis ianceoiata and 100-fold diluted Cornus officinalis were 25% and 73%, respectively. The addition of salt to Codonopsis lanceolata and Cornus officinalis slightly decreased their ${\alpha}$-glucosidase inhibitory activities. According to the addition of Cornus officinalis to Codonopsis lanceoiata, the ${\alpha}$-glucosidase inhibitory activities of the resulting mixture were highly increased. We anticipate that these results will be used as basic data for the development of new bifunctional foods.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.38 no.1
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• pp.1-5
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• 2005

This study investigated and compared chemical properties and biological activities of four commercial sikhae products (three flat-fish, Pleuronectes herzensteini, Sikhae: product A, B, C; one Alaska pollack, Theragra chalcogramma, Sikhae: product D) and Alaska pollack Sikhae (product E). Total acidity, amino nitrogen, salinity and pH in all products were in ranges of $0.47-1.93\;g\%$, $145.94-204.81\;mg\%$, $3.40-4.00\%$ and 4.38-5.55, respectively. All products showed antimicrobial activities against Bacillus ceraus subtilis, B. cereus, Staphylococcus aureus, Listeria monocytogenes, Vibrio parahaemolyticus, except for Salmonella typhimurium. Particularly the activities were more effective with gram positive bacteria than gram negative bacteria. Antioxidative $(EDA_{50};\;14.20\;mg/mL)$ and Fibrinolytic (0.95 plasmin unit/mL) activities of product E were much stronger than commercial Sikhae products (EDA50; 18.87-34.60 mg/mL and 0.69-0.85 plasmin unit/mL).

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.655-659
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• 1998

Bacterial strain showing the strong fibrinolytic activity (2.04 plasmin unit) was screened from Jeot-Gal, Korean salt-fermented fish collected from various region. For the identification, when the strain was characterized morphologically, culturally, and biochemically, it was identified to Bacillus pumilus. And, when the fatty acids composition of the strain was analyzed, it was identified to Bacillus atropheus. Finally, the 16S rRNA partial sequence (V3 region) showed that the fibrinolytic stain screened from Jeot-Gal was identified as Bacillus subtilis. So, we named it Bacillus subtilis KJ-48.

• Korean Journal of Medicinal Crop Science
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• v.14 no.4
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• pp.195-201
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• 2006

The fibrinolytic activities of soluble proteins extracted from young leaves of Camellia japonica L. were studied. Fibrinolvity activity of extract from partitions of C. japonica L. showed 1.6-2.0 times higher than plasmin used as positive control. The fibrinolytic enzyme was confirmed directly from young leaves of C. japonica L. by a fibrin Plate and fibrin zymography. The protein was composed of a single polypeptide and its apparent molecular weight was found to be 45 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for the fibrinolytic activity were pH 5.5 and $30^{\circ}C$, respectively. Also, the fibrinolytic activity was clearly inhibited by PMSF and TLCK, suggesting that it is a member of the trypsin-like serine protease. All these results suggest the protease is a fibrinolytic enzyme belong to a family of trypsin-like serine protease.

• Journal of Life Science
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• v.20 no.6
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• pp.838-844
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• 2010

A fibrinolytic enzyme of Streptomyces corcohrussi from soil sediment was purified by chromatography using DEAE-Sephadex A-50 and Sephadex G-50. The analysis of SDS-polyacrylamide gel suggested that the purified enzyme is a homogeneous protein and the molecular mass is approximately 34 kDa. The purified enzyme showed activity of 0.8 U/ml in a plasminogen-rich fibrin plate, while its activity in a plasminogen-free fibrin plate was only 0.36 U/ml. These results suggested that the purified enzyme acts as a plasminogen activator. The fibrinolytic activity of the enzyme under the supplementation of protease inhibitors, $\varepsilon$-ACA, t-AMCHA and mercuric chloride in the enzyme reaction was less than 24%, indicating that it could be modulated by the plasmin and/or fibrinogen inhibitors involved in the fibrinogen-to-fibrin converting process. As time passed, $Zn^{2+}$, a heavy metal ion, inhibited the activity to 34.1%. The optimum temperature of the purified enzyme was approximately $50^{\circ}C$ and over 92% of the enzyme activity was maintained between pH 5.0 and 8.0. Therefore, our results provide a potential fibrinolytic enzyme as a noble thrombolytic agent from S. corcohrussi.