• Title/Summary/Keyword: degree of binding

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Protection of Metal Stress in Saccharomyces cerevisiae: Cadmium Tolerance Requies the Presence if Two ATP-Binding Domains of Hsp 104 Protein

  • Lee, Gyeong Hui;Eom, Jeong Hun
    • Bulletin of the Korean Chemical Society
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    • v.22 no.5
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    • pp.514-518
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    • 2001
  • We have explored the importance of two ATP binding domains of Hsp104 protein in protection of yeast cells from cadmium exposure. In the previous study we have discovered that the presence of two ATP binding sites was essential in providing heat sh ock protection as well as rescuing cells from oxidative stress. In this paper we first report wild type cell with functional hsp104 gene is more resistant to cadmium stress than hsp104-deleted mutant cell, judging from decrease in survival rates as a result of cadmium exposure. In order to demonstrate functional role of two ATP binding sites in cadmium defense, we have transformed both wild type (SP1) and hyperactivated ras mutant (IR2.5) strains with several plasmids differing in the presence of ATP binding sites. When an extra copy of functional hsp104 gene with both ATP binding sites was overexpressed with GPD-promoter, cells showed increased survival rate against cadmium stress than mutants with ATP binding sites changed. The degree of protection in the presence of two ATP binding sites was similarly observed in ira2-deleted hyperactivated ras mutant, which was more sensitive to oxidative stress than wild type cell. We have concluded that the greater sensitivity to cadmium stress in the absence of two ATP binding sites is attributed to the higher concentration of reactive oxygen species (ROS) produced by cadmium exposure based on the fluorescence tests. These findings, taken all together, imply that the mechanism by which cadmium put forth toxic effects may be closely associated with the oxidative stress, which is regulated independently of the Ras-cAMP pathway. Our study provides a better understanding of cadmium defense itself and cross-talks between oxidative stress and metal stress, which can be applied to control human diseases due to similar toxic environments.

Effect of Mugwort Powder on Physicochemical Properties, Paste and Gel of Rice Flour (쑥가루를 첨가한 습식 및 건식 쌀가루의 이화학적 특성)

  • 김영인
    • Korean Journal of Rural Living Science
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    • v.8 no.2
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    • pp.125-129
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    • 1997
  • The effect of mugwort powder on some physicochemical properties of rice flour, the viscosity of rice flour pastes and the degree of gelatinization of pastes and gels were investigated. By addition of mugwort, water binding capacity, swelling power, and solubility of rice flour were increased, also the viscosity and, the degree of gelatinization of rice flour pastes and gels were increased. Generally wet rice flour-mugwort pastes and gels were better than those of dry rice flour-mugwort. And viscosity and gelatinization of the rice-mugwort pastes and gels were decreased slightly during storage at $20^{\circ}C$ for 7 days.

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Comparative Study on the Degumming Methods of Hemp Fiber (대마섬유의 정련 방법에 관한 비교 연구)

  • Lim, Hyeong-Gyu;Kim, Hee-Sook
    • Fashion & Textile Research Journal
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    • v.22 no.4
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    • pp.523-533
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    • 2020
  • Previous studies are used to examine cellulose content, degumming period, fiber quality, production yield, production cost, development limit of fiber according to physical, chemical, and microbial degumming methods. Three types of degumming methods are used to measure surface condition after degumming, necessity of additional degumming and degree of impurity removal. First, previous studies confirmed that the microbial degumming method is superior in terms of cellulose content, fiber quality, production yield, production cost, and fiber development possibility. Second, surface condition and the necessity of additional degumming were analyzed by SEM. The black skin binding material was removed in the case of the Sangnangyi and chemical degumming; however, it was insufficient and further degumming was required. Skin fiber binding material was removed in the case of microbial degumming and the surface was cleanest after degumming; in addition, most showed the form of yarn decomposition. The FT-IR spectrum determined the degree of removal of impurities and showed that it can utilize inherent physical properties as the best degumming method. The degree of removal of pectin and lignin by microbial degumming was cleanest with hemicellulose also reduced by microbial degumming.

The Distribution of Estrogen Receptor in Various Organs of Rabbit (토끼 각장기의 Estrogen 수용체의 분포)

  • Son, Ho-Young;In, Jae-Whan;Min, Byong-Sok
    • The Korean Journal of Nuclear Medicine
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    • v.12 no.2
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    • pp.1-5
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    • 1978
  • For clinical application of radioreceptor assay, we studied preliminarily the distribution of estrogen receptor in various organs of rabbit by a dextran-charcoal method using $6,7-^3H-estradiol$. The results were expressed as binding index, which is the ratio of specific estradiol receptor binding radioactivity to total radioactivity. The materials consist of 5 female rabbits and 3 male rabbits. The results were as follows: 1) Female rabbits The binding index was highest in the uterine tissue. This binding index of the uterine tissue was 9.4 times that of the liver, 21.9 times that of the kidney, 24.6 times that of the brain, 28.1 times that of the lung and 65.7 times that of the muscle. 2) Male rabbits The binding index was highest in the liver and decreased in the order of the kidney, the testis, the lung, the brain and the muscle. It is suggested that the estrogen receptor is not confined to any specific target organ but is widely distributed in the various organs, to a different degree.

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Drug-biomacromolecule interaction IV

  • Kim, Chong-Kook;Yang, Ji-Sun;Lim, Yun-Su
    • Archives of Pharmacal Research
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    • v.6 no.1
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    • pp.55-62
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    • 1983
  • Binding of six cephalosporins (cefotaxime, cefuroxime, cefazoline, cephalothin, cephaloridine, cephacetrile) to bovine serum albumin was studied. Fluorescence probe technique and difference spectrophotometry were employed to evaluate the nature and degree of association of cephalosporin albumin complex. 1-Anilinonaphthalene-8-sulfonate (ANS) was used as the fluorescence probe. 2-(4'-hydroxybenzeneazo) benzoic acid(HBAB) was employed as the UV spectrophotometric probe. Compentitive bindings between cephalosporins and probes were observed. The number of binding sites of bovine serum albumin for each cephalcsporin is 2. Among six cephaloporins, cefotaxime has the highest binding constant followed by cafazoline, cefuroxime, cephalothin, cephaloridine and cephacetrile.

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Separation of Calcium-binding Protein Derived from Enzymatic Hydrolysates of Cheese Whey Protein

  • Kim, S.B.;Shin, H.S.;Lim, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.712-718
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    • 2004
  • This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

Effect of Pressure on Catalytic Properties of Glutamate Racemase from Aquifex pyrophilus, an Extremophilic Bacteria

  • Lee, Ki-Seog;Chi, Young-Min;Yu, Yeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.149-152
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    • 2002
  • The effect of pressure on the catalytic properties of glutamate racemase from Aquifex pyrophilus, an extremophilic bacterium, was investigated. The activation volume for the overall reaction $({\Delta}V^{\neq})$ and catalysis $({{Delta}V_{cat}}^{\neq})$ was -96.97 ml/mol and 4.97 ml/mol, respectively, while the reaction volume for the substrate binding (${\Delta}V_{K_m^-1}$) was -101.94 ml/mol. The large negative ${\Delta}V^{\neq}$ for the overall reaction indicated that the pressurization of glutamate racemase resulted in enhanced catalytic efficiencies. In addition, this value was also due to the large negative ${Delta}V_{K_m^-1}$ for the substrate binding. The negative value of ${Delta}V_{K_m^-1}$ implied that the conformational changes in the enzyme molecule occurred during the substrate binding process, thereby increasing the degree of hydration. The small value of ${{Delta}V_{cat}}^{\neq}$suggested that the pressure did not affect the glutamate racemase catalysis after the substrate binding.

Percutaneous Absorption of Recombinant h-EGF through Normal, Stripped and First-Degree Burn Skin (상피세포 성장인자의 경피흡수: 정상피부, 각질제거피부 및 화상피부에 있어서)

  • Cho, Ae-Ri;Lee, Jung-Uk;Ahn, Byung-Lak;Chung, Joo-Young;Koh, Yeo-Wook;Shim, Chang-Koo
    • Journal of Pharmaceutical Investigation
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    • v.26 no.1
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    • pp.29-32
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    • 1996
  • In vivo and in vitro skin permeation of $recombinant^{125}$ I-EGF through normal, stripped and the first degree burn skin were studied. The in vitro skin permeation rate through the first degree burn skin $(296\;cpm/cm^2/hr)$ and the stripped skin $(1131\;cpm/cm^2/hr)$ were 3.5 times and 13 times higher, respectively, as compared with the one through normal skin. In vivo absorption study with the first degree burn skin, the peak concentration of EGF in the skin was achieved at 1-3 hr and decreased afterward up to 8 hr with an elimination constant of $1.31{\times}10^{-3}\;g/ml/hr$. To investigate the higher elimination rate of EGF in burn skin, binding and metabolism studies were conducted. No significant metabolism of EGF in burn skin $(100^{\circ}C,\;5-second\;burning)$ was observed. With the presence or unlabelled-EGF $^{125}I-EGF$ permeation through the burn skin showed higher permeation rate than the one without unlabelled-EGF. The result nay indicate that EGF-receptor binding play a role in determining the skin permeation rate.

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Effects of Individual Fatty Acids on Receptor-Mediated Binding, Internalization and Degradation of $[^{125}I]LDL$

  • Choue, Ryo-Won;Cho, Byung-Hee Simon
    • BMB Reports
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    • v.30 no.1
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    • pp.1-6
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    • 1997
  • The ability of Hep-G2 cells to process $[^{125}I]LDL$ under basal conditions was investigated. The receptor-binding and internalization of $[^{125}I]LDL$ increased with the time of incubation in a saturable manner. After 4 h of incubation, 31.4 ng of $[^{125}I]LDL$ was cell bound. The cells rapidly internalized $[^{125}I]LDL$ via specific, receptor-mediated endocytosis. The amount of internalized $[^{125}I]LDL$ reached a maximun of 96.7 ng at 2 h of incubation and remained constant for the next 2 h. The rate of degradation of internalized $[^{125}I]LDL$ proceeded in a linear manner over the entire 4 h of incubation after an initial lag period. The effects of individial fatty acids (C18:0. C18:1, C18:2. and C18:3), differing in their degree of unsaturation. on the receptor-binding, internalization and degradation of $[^{125}I]LDL$ were also investigated. Inclusion of 1.0 mM of each fatty acid into the culture medium significantly increased $[^{125}I]LDL$ metabolism in Hep-G2 cells. Among the fatty acids tested, stearic acid had the least effect on the receptor-binding activity. There were no significant differences among the unsaturated fatty acids in LDL-receptor binding. The effect of individual fatty acids on the $[^{125}I]LDL$ uptake was similar to that of the receptor-binding. showing a significantly lower effect with stearic acid. The amount of degraded material of internalized $[^{125}I]LDL$ was the lowest with stearic acid when it was compared with unsaturated fatty acids.

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Isolation of Iron and Calcium-Binding Peptides from Cottonseed Meal Protein Hydrolysates (면실박 단백질로부터 가수분해물 제조 및 철분, 칼슘 결합 펩타이드의 분리)

  • Choi, Dong-Won;Kim, Nam-Ho;Song, Kyung Bin
    • Journal of Applied Biological Chemistry
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    • v.55 no.4
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    • pp.263-266
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    • 2012
  • Isolation of iron and calcium-binding peptides derived from cottonseed meal protein (CMP) hydrolysates was investigated. The degree of hydrolysis of CMP by Flavourzyme was monitored using trinitrobenzenesulfonic acid method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Enzymatic hydrolysis of CMP for 12 h was sufficient for the preparation of CMP hydrolysates, and the hydrolysates were membrane-filtered under 3 kDa as a molecular weight. The filtered solution was fractionated using Q-Sepharose fast flow, Sephadex G-15, and reversed phase-high performance liquid chromatography for iron and calcium-binding peptides. As a result, F51 fraction was obtained as the best candidate for calcium and iron chelation, and the isolated iron and calcium-binding peptides can be used as functional food additives, similar to iron and calcium supplements.