• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2004.10a
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• pp.93-93
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• 2004

• Biomedical Science Letters
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• v.3 no.2
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• pp.89-94
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• 1997

The authors characterized the proteins of the crude antigen obtained from Clonorchis sinensis worm and excretory-secretory and billis from rabbits, experimentally infected for 3 months. Protein composition was observed after adding a cysteine proteinase inhibitor E-64 and a serine proteinase inhibitor PMSF, respectively. SDS-PAGE of the crude antigen from C. sinensis recovered from the infected rabbits, the crude antigen from the adult worm excretory-secretory, and the crude antigen from billis of the rabbits resolved 26, 27 and 19 profiles between 200-9 kDa, respectively. When E-64 supplemented 29, and 22 bands, respectively. More study should be carried out in the future on the immunological characteristics and the monoclonal antibody of the each antigen.

• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.90-95
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• 2001

Kinetic Analysis of Cathepsin B Inhibitor Using a Spectrophotometric Assay. Han, Kil-Hwan and SangDal Kim*. Department of Applied MicrobioJ0f5Yt Yeungnam UniversitYt Kyongsan 77 2-749, Korea - The KHS 10, C4Hl10~6 formula produced from Streptomyces luteogriseus KT-] 0 effectively inhibited a lysosomal cysteine proteinase, cathepsin B. It inhibited the enzyme activity of cathepsin B competitively when the N a-CBZ-Llysine p-nitrophenyl ester HC] (CLN) was used as a substrate. The inhibition const:mt (Ki) of KHS 1 0 for cathepsin B detennined by spectrophotometeric assay was 430 nM. The effective inhibition of cathepsin B was observed at $25^{\circ}C$ :md pH 6.0. The cathepsin B inhibitor, KHSlO needed a preincubation of cathepsin B with the inhibitor for over 5 min. The KHS 10 preserved over 80% inhibition activity even after heat-treatment at $100^{\circ}C$ for ] hr.

• Biomolecules & Therapeutics
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• v.24 no.3
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• pp.244-251
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• 2016

Understanding the developmental mechanisms of humoral immunity against intranasal antigens is essential for the development of therapeutic approaches against air-borne pathogens as well as allergen-induced pulmonary inflammation. Follicular helper T (Tfh) cells expressing CXCR5 are required for humoral immunity by providing IL-21 and ICOS costimulation to activated B cells. However, the regulation of Tfh cell responses against intranasal antigens remains unclear. Here, we found that the generation of Tfh cells and germinal center B cells in the bronchial lymph node against intranasal proteinase antigens was independent of $TGF-{\beta}$. In contrast, administration of STAT3 inhibitor STA-21 suppressed the generation of Tfh cells and germinal center B cells. Compared with wild-type OT-II T cells, STAT3-deficient OT-II T cells transferred into recipients lacking T cells not only showed significantly reduced frequency Tfh cells, but also induced diminished IgG as well as IgE specific for the intranasal antigens. Cotransfer study of wild-type OT-II and STAT3-deficient OT-II T cells revealed that the latter failed to differentiate into Tfh cells. These findings demonstrate that T cell-intrinsic STAT3 is required for the generation of Tfh cells to intranasal antigens and that targeting STAT3 might be an effective approach to ameliorate antibody-mediated pathology in the lung.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.8
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• pp.1100-1105
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• 2006

During involution of the mammary gland after the lactation period, the gland undergoes an extensive epithelial cell death. In our previous study, overexpression of an extracellular proteinase inhibitor (Expi) gene accelerated apoptosis of mammary epithelial cells. Here we found that expression of the cathepsin D gene was induced in the Expi-overexpressed apoptotic cells. To understand the role of cathepsin D in apoptosis, we transfected cathepsin D gene into mammary epithelial HC11 cells and established the stable cell lines overexpressing the cathepsin D gene. We found that overexpression of the cathepsin D gene partially induced apoptosis of mammary epithelial cells. Expression patterns of the cathepsin D gene were examined in mouse mammary gland at various reproductive stages. Expression of the cathepsin D gene was increased during involution stages compared to lactation stages, and highest expression levels were shown at involution on day 4. We also examined expression of the cathepsin D gene in various mouse tissues. Mammary gland at involution on day 2 showed highest levels of cathepsin D mRNA of the mouse tissues that we examined. Liver tissues showed high levels of cathepsin D expression. These results demonstrate that cathepsin D may contribute to the apoptotic process of mammary epithelial cells.

• Journal of Animal Science and Technology
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• v.45 no.2
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• pp.319-326
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• 2003

Angiotensin-I converting enzyme(ACE)inhibitor was isolated from beef by-products. The beef by- product hydrolysates prepared with various proteases were tested for the inhibitory effects against ACE. The proteases used were proteinase A from bakers yeast, protease type ⅩIII fungal and thermolysin. The maximum inhibitory effect was observed after hydrolysis for 12hrs(beef heart) and 24hrs(beef spleen), respectively. After gel filtration, IC50 value was 0.37mg/ml in beef heart and 1.84mg/ml in beef spleen. After RP-HPLC, the IC50 value of peak 1, peak 2, peak 3 and peak-4 were 0.28mg/ml, 0.26mg/ml, 0.25mg/ml and 0.35mg/ml, respectively. In the results of amino acid composition of peak 1, peak 2, peak 3 and peak 4, it was observed that peak 1 was consisted mainly of glycine and methionine, peak 2 was proline, cystine and methionine, peak 3 was proline and peak 4 was alanine, methionine and leucine. In conclusion, beef heart hydrolysate treated with thermolysin+ proteinase A was shown to have the highest inhibitory effect for 12hrs incubation at 37$^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.145-153
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• 1986

For the inactivation of venoms, the chemical methods are generally applied. In the chemical method many works have been carried out with the chemical reagents and immunological antiserums. However, all inhibitory effect of these chemicals acting on snake venomes may well be due not to the specific, but to the nonspecific inhibitory action. Therefore, it is necessary to separate venom into its compositional active proteins and develop specific inhibitor which acts on the each protein. Until now, there have not been any reports about the substance which acts on snake venom as a specific inhibitor. Recently in 1979, we had actually isolated a specific venom inhibitor(ISV) which has a strong inhibitory activity against the proteinase of snake venom of Colubridae. In our experiments described here, a strain of Aspergillus sp., isolated from soil, was able to produce a biological active substance. The partial crystallized substance had a strong inhibitory activity against hemorrhagic action of snake venom of Colubridae. For the inhibitory action of the sample on the lethality of venom, the substance prevented completely the lethal action of the hemorrhagic factor when they were treated with enough amount of the substance. The edema factor of whole venom of Agristrodon bromohoffi brevicaudus was completely inhibited, but those of HR-I and HR-II of Trimeresurus flavoviridis venom were inhibited about 50%, when they were treated with the substance of half amount of venom. On the other hand, from the result of subcutaneous hemorrhage in a rabbit, it was concluded that two kinds of antihemorrhagic substance might be produced by the strain used in this work.

• Applied Biological Chemistry
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• v.45 no.4
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• pp.212-217
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• 2002

We purified polyphenols from persimmon leaf and tested their biological activity. The 60% acetone extract was lyophilized and applied to test enzyme inhibition of glucosyltransferase and tyrosinase. GTase was 82.4% inhibited at $1.8{\times}10^{-1}$ mg/ml and tyrosinase 21.7% inhibited at 0.8 mg/ml. The acetone extract was fractionated into F-1, 2, 3, 4, 5 by Sephadex Q-50 gel filtration and the fraction-1 and 2 showed higher enzyme inhibition activity than the other fractions. To the Proteinase K treatment and autoclaving of the two fractions had no effect on the enzyme activity, but these results suggested that active fraction was not protein but phenol ring completed compounds. By Sephadex LH-20, MCI-gel and Bondapak $C_{18}$ column chromatographies, compouds 1, 2, 3 and 4 from F-1 fraction, compounds 5 and 6 from F-2 fraction and compounds 7 , 8 from F-3 fraction were purified and re-crystallized. The purified compounds was assumed to be condensed tannins of frame flavan-3-ol frame on the basis of color reagent reaction and to be a mixture of monomer, dimer and trimer according to TLC analysis.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.425-431
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• 1996

Human neutrophil elastases (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, are regulated by plasma proteinase inhibitors, alpha-proteinase inhibitor and ${\alpha}_2-macroglobulin$. Under certain pathological conditions, however, released enzymes or abnormal function of inhibitors may cause various inflammatory disease. NSAIDs have been clinically applied for treatment of inflammatory diseases. Inhibition of cyclooxygenase is a known mechanism of action of NSAIDs in the treatment of inflammatory disease. In in vitro experiments, HNElastase was inhibited by naproxen, phenylbutazone, and oxyphenbutazone, but ibuprofen, ketoprofen, aspirin, salicylic acid, and tolmetin did not inhibit elastase. HNElastase was also inhibited by chelating agents, EDTA & EGTA, and tetracyclines. Removal of divalent metal ions by EDTA caused inhibition of elastase, and reconstitution of the metal ions recovered the enzyme activity to a certain level. Frequencies and contours in the Raman spectra of various conditions of human neutrophil elastase undergo drastic changes upon partial removal and/or reconstitution of calcium and zinc ions. The metal ion content dependent activities and change of the contour of the Raman spectrogram suggest us that the mechanism of action of a chelator or chelator-like agents on neutrophil elastase may be related to the conformational change at/or near the active site, especially -C=O radical or -COOH radical.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.17 no.3
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• pp.235-244
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• 2007

By comparing the proteins from the workers exposed to styrene with the ones from controls, it may be possible to identify proteins that play a role in the occurrence and progress of occupational disease and thus to study the molecular mechanisms of occupational disease. In order to find the biomarkers for assessing the styrene effects early, before clinical symptoms develop and to understand the mechanisms of adverse health effects, we surveyed 134 employees, among whom 52 workers(30 male and 22 female) were chronically exposed to styrene in 10 glass-reinforced plastic boat manufacturing factories in Korea and 82 controls had never been occupationally exposed to hazardous chemicals including styrene. The age and drinking habits and serum biochemistry such as total protein, BUN and serum creatinine in both groups were significantly different. Exposed workers were divided into three groups according to exposure levels of styrene(G1, below 1/2 TLV; G2, 1/2 TLV to TLV; G3, above TLV). The mean concentration of airborne styrene in G1 group was $10.93{\pm}11.33ppm$, and those of urinary mandelic acid(MA) and phenylglyoxylic acid(PGA) were $0.17{\pm}0.21$ and $0.13{\pm}0.11g/g$ creatinine, respectively. The mean concentration of airborne styrene in G2 and G3 groups were $47.54{\pm}22.43$ and $65.33{\pm}33.47ppm$, respectively, and levels of urinary metabolites such as MA and PGA increased considerably as expected with the increase in exposure level of styrene. The airborne styrene concentration were significantly correlated to the urinary concentration of MA(r=0.784, p=0.000) and PGA(r=0.626, p<0.001). In the 2D electrophoresis, the concentration of five proteins including complement C3 precursor, alpha-1-antitrypsin(AAT), vitamin D binding protein precursor(DBP), alpha-1-B-glycoprotein(A1BG) and inter alpha trypsin inhibitor(ITI) heavy chain-related protein were significantly altered in workers exposed to styrene compared with controls. While expression of complement C3 precursor and AAT increased by exposure to styrene, expression of DBP, A1BG and ITI heavy chain-related protein decreased. These results suggest that the exposure of styrene might affects levels of plasma proteinase, carriers of endogenous substances and immune system. In particular, increasing of AAT with the increase in exposure level of styrene can explain the tissue damage and inflammation by the imbalance of proteinase/antiproteinase and decrease of DBP, A1BG and ITI heavy chain-related protein in workers exposed to styrene is associated with dysfunction and/or declination in immune system and signal transduction