• BMB Reports
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• v.29 no.6
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• pp.555-563
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• 1996

A membrane-bound phosphatidylinositol 4-kinase (PI 4-kinase) was separated in a sucrose gradient and solubilized with 1% Triton X-100 from mouse brain. The enzyme was purified 2,952-fold by various chromatographic techniques including DEAE-cellulose, PI-Sepharose and Sephacryl S-200 gel filtration. The molecular weight of PI 4-kinase was approximately 76 kDa by gel filtration and 70.8 kDa by SDS-polyacrylamide gel electrophoresis. The purified enzyme exhibited specific activity of 11.2 nmol/min/mg protein and pi value of 4.7. Kinetic analysis of the PI 4-kinase indicated apparent $K_m$, values of 190 ${\mu}M$ and 120 ${\mu}M$ for phosphatidylinositol and ATP, respectively. The maximal activity of this purified enzyme was observed at pH 7.4 at an incubation temperature of $37^{\circ}C$. The enzyme activity was significantly activated by $Mg^{2+}$, $Mn^{2+}$ and $Fe^{2+}$, and inhibited severely by $Ca^{2+}$. PI 4-kinase was proved to be pure in its immunoblot test by polyclonal antibody prepared from immunized rabbit sera. By this test, we were able to detect the existence of the same type of PI 4-kinase from other mouse organ tissues, such as liver, heart, kidney and spleen. Furthermore, similar immunoblot analysis with the same antisera recognized the different epitopes of PI 4-kinase proteins from various organs of rabbit, chinese hamster and rat.

• Journal of Ginseng Research
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• v.42 no.3
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• pp.343-351
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• 2018

Background: Red ginseng is a popularly used traditional medicine with antiaging effects in Asian countries. The present study aimed to explore the changes in protein expression underlying the mechanisms of life span extension and antiaging caused by red ginseng extract (RGE) in Drosophila melanogaster. Methods: A proteomic approach of two-dimensional polyacrylamide gel electrophoresis (2-DE) was used to identify the differential abundance of possible target proteins of RGE in D. melanogaster. The reliability of the 2-DE results was confirmed via Western blotting to measure the expression levels of selected proteins. Proteins altered at the expression level after RGE treatment (1 mg/mL) were identified by matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry and by searching against the National Center for Biotechnology nonredundant and Uniprot protein databases. The differentially expressed proteins were analyzed using bioinformatics methods. Results: The average survival life span of D. melanogaster was significantly extended by 12.60% with RGE treatment (1 mg/mL) compared to untreated flies. This followed increased superoxide dismutase level and decreased methane dicarboxylic aldehyde content. Based on the searching strategy, 23 differentially expressed proteins were identified (16 up-regulated and 7 down-regulated) in the RGE-treated D. melanogaster. Transduction pathways were identified using the Kyoto Encyclopedia of Genes and Genomes database, and included the hippo and oxidative phosphorylation pathways that play important roles in life span extension and antiaging process of D. melanogaster. Conclusion: Treatment with RGE in D. melanogaster demonstrated that mechanisms of life span extension and antiaging are regulated by multiple factors and complicated signal pathways.

• Korean Journal of Air-Conditioning and Refrigeration Engineering
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• v.2 no.1
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• pp.37-48
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• 1990

The drag and heat transfer reduction phenomena and degradation effects of drag reducing polymer solutions which are known as the viscoelastic fluids are investigated experimentally for the turbulent circular tube flows. Two stainless steel tubes are used for the experimental flow loops. Aqueous solutions of Polyacrylamide Separan AP-273 with concentrations from 300 to 1000 wppm are used as working fluids. Flow loops are set up to measure the friction factors and heat transfer coefficients of test tubes in the once-through system and the recirculating flow system. Test tubes are heated by power supply directly to apply constant heat flux boundary conditions on the wall. Capillary tube viscometer and falling ball viscometer are used to measure the viscous characteristics of fluids and the characteristic relaxation time of a fluid is determined by the Powell-Eyring model. The order of magnidude of the thermal entrance length of a drag reducing polymer solution is close to the order of magnitude of the laminar entrance length of Newtonian fluids. Dimensionless heat transfer coefficients of the viscoelastic non-Newtonian fluids may be represented as a function of flow behavior index n and newly defined viscoelastic Graetz number. As degradation continues viscosity and the characteristic relaxation time of the testing fluids decrease and heat transfer coefficients increase. The characteristic relaxation time is used to define the Weissenberg number and variations of friction factors and heat transfer coefficients due to degradation are presented in terms of the Weissenberg number.

• Journal of Oral Medicine and Pain
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• v.22 no.1
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• pp.65-80
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• 1997

Cytochrome P450 is an oxidase involved in oxidation of alcohol and is known to be an activator of carcinogen. The present study was perfomed to analyze the effect of diabetes and cold stress on the expression of Cytochrome P450 IIE1(CYPIIE1) in the liver and salivary glands in rats by an immunoblot analysis. Sixty three divided into 4 groups; 1) 20 rats belonging to group I were allowed diabetes (40mg/kg. I.V.) 2) 20 rats of group II were bathed in cold water for 30 seconds twice a day 3) 20 rats comprising group III were received diabetes and cold stress as described above 4) 3 rats of group IV were selected as a control. The rats were sacrificed at the end of the same day 1, 2, 3, 4 weeks experiment. The liver and parotid glands were removed and stored at $-70^{\circ}C$ until use. The stored organs were homogenized for 10 seconds and the supernatants were obtained by centrifugation. The proteins of the supernatants were separated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis and subjected to Western blotting. The blotted membranes were incubated with polyclonal antibodies to CYPIIE1. And sepimens were observed with light microscope also under the Hematoxillin-Eosin staining. The obtained results were as follows : 1. In diabetes group, acini had changed to degeneration severely 1 week experiment, but repaired gradually in lapse of time. 2. In diabetes group, septal connective tissue had changed to degeneration little by little from 1 week after experiment, and progressed severely in lapse of time. 3. In stress group, acini had not changed remarkably, but slightly separated each other 3 weeks after experiment. 4. In diabetes and stress group, histological feature had changed remarkably campare with in the group of diabetes only. 5. In all experimental group, CTPIIE1 had expressed remarkably in the liver tissue, but not in the parotid gland tissues. 6. In diabetes and stress group, CYPIIE1 had expressed remarkably campare with in the group of diabetes only.

• 보존과학연구
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• s.20
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• pp.5-20
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• 1999

We have analyzed the allele and genotype frequencies from 10 fractions of ancient human skeleton in 3 pieces of Jar coffin excavated from Naju Bokamni3rd tumulus by PCR amplification, high resolution polyacrylamide gelelectorphoresis and silver staining. We could isolate human genomic DNA from 3 bone fractions but the rest of them could not be used as materials due to being decayed. We could detect sex determination as male and 3 genotypes of STR system, HUMTHO1, HUMTPOX and HUMC5F1PO from the bone fraction of left side in Jar coffin 3 and see the slightly reaction suggesting the sex as male from the bone fraction of the left side in Jar coffin 2 and female from the right side in Jar coffin 3.We have also analyzed the genotype frequencies of mitochondria from the bone fractions of the left side and the right side in jar coffin 3, respectively. From the result of indetifiying at nucletide position between 16018 and 16378of the base of hyper variable region(HV1) in the control region, We can presume that the both bones have the same maternal inheritance.

• Korean Journal of Plant Resources
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• v.12 no.3
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• pp.198-203
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• 1999

7S and 11S globulins are two major storage proteins in soybean seed. For improving the quality of soybean seed protein, an increase of 11S/7S ratio would be a desirable objective because 11S globulin contains much more sulfur-containing amino acids than 7S globulin. In this study, six soybean varieties grown at three locations were used for genetic variation analysis of 7S and 11S globulins. It was possible to screen the soybean genotypes having aberrant subunit compositions of the two globulins by a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). So, heritabilities, genotypic and phenotypic correlations among eight globulin fraction contents of soybean seeds were estimated. The mean value of 7S and 11S globulin fraction contents were 38.9% and 61.1%, respectively, and the ratio of 7S to 11S globulin ranged from 0.58 to 0.74. The high heritability value was found in $\beta$ subunits but the values of acidic and basic subunits were relatively low. Genotypic correlations were higher than the corresponding phenotypic correlations in most of globulin subunit contents. $\beta$ subunits was negatively correlated with $\alpha$ and $\alpha$' subunits among 7S fractions, while no significant correlation between $\alpha$ and $\alpha$' subunits could be found In case of 11S fractions, acidic and basic subunits exhibited no genotypic but negative phenotypic correlation.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.816-822
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• 2005

Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

• Korean Journal of Food Science and Technology
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• v.35 no.2
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• pp.260-265
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• 2003

Transglutaminase (TG) was prepared from Streptomyces mobaraensis to improve texture and self-life of food. In preliminary experiments, texture of the dough was not improved due to the interference in microbial TG reaction by proteases present in the crude enzyme. Among the cation exchange resins tested for the removal of proteases, MonoPlus S 100 was the most efficient. Further purification steps with a quaternary ammonia salt resin and gel permeation chromatography effectively removed proteases from crude enzyme. Molecular weight of purified enzyme was about 38,000 on SDS-polyacrylamide gel electrophoresis. Farinograph data showed the addition of purified enzyme to wheat flour gave higher stability and lower weakness values those that of crude enzyme.

• Korean Journal of Food Science and Technology
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• v.17 no.5
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• pp.377-388
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• 1985

The changes in protein fractions and its gel electrophoretic pattern of rapeseeds (Brassica napus L) were investigated during germination at $25^{\circ}C$ under dark condition. The major protein fraction was found to be albumin 25.0%) and globulin (24.6%). Both fractions were decreased throughout germination, particularly significant for albumin, while prolamine(2.2%) and glutelin (1.8%) showed an initial decrease followed by a slow increase at the later stage of germination. The initial 5-6 peaks of gel electrophoresis were reduced to a few after 45 hours. The absorption spectrum at the range of 400-700 nm showed a significant increase in absorbance for sodium hexametaphosphate (SHMP) extract of rapeseeds. The protein extractability with SHMP was not significantly affected by germination.

• Korean Journal of Food Science and Technology
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• v.20 no.2
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• pp.263-271
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• 1988

Fatty acid composition and electrophoretic patterns of protein of Korean soybeans were studied, The protein content showed an inverse relation with the lipid content, The content of lipid types of total lipid was in the order of neutral lipid, glycolipid and phospholipid, Cultivar 102-B had higher content of neutral lipid but lower content of phospholipid compared with others. The main component of neutral lipid was triglyceride followed by sterol. The main fatty acids of total lipid were linoleic, oleic and palmitic acids, which comprised over 85% of total fatty acids. The main fatty acid of neutral lipid, glycolipid and phospholipid were linoleic acid. The polyacrylamide gel electrophoretic pattern of protein showed seven bands. The cultivar 102-B had the highest content of IIS globulin. Sodium-dodecyl sulfate PAG electrophoresis demonstrated that all soybeans had similar composition of subunit.