• Title/Summary/Keyword: protein fraction

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Evidence for Regulation of Interaction of Endogenous Protein Kinase C(Pkc) Substrates with Plasma Membrane by PKC Down-Regulation in K562 Cells

  • Kim, Young-Sook
    • Archives of Pharmacal Research
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    • v.18 no.5
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    • pp.301-307
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    • 1995
  • In the particulate fraction obtained from PKC-down regulated K562 cells by treatment for 24 h with 200nM TPA, phosphorylation of two proteins with molecular weight, 100 kDa and 23 kDa (designated p100 and p23, respectvely) was depleted and addition of exogenous purified PKC to this fraction failed to testore their phosphorylation. However, in the soluble fraction, all of phosphoproteins abolished by long-term treatment with TPA were restored by exogenously added PKC. Phosphorylation of two proteins was increased by short-term tretment (20 min), and diminished with the persistant exposure to TPA as well as at a concentration as low as 1nM. When K562 cells were treated with 1 nM and 200 nM TPA for 24 h, phosphorylation of p100 was restored with or without exogenous PKC on 2-3day and 6day after removal of treated TPA, respectively. Two-dimensional electrophoresis of phosphoproteins revealed that phosphorylated p100 (pl=5.9) and p66 species were completely absent from the particulate fraction of K562 cells treated with 200nM TPA for 24 h. These results suggest that the interaction of sensitive endogenous substrates, p100 and p23 with the plasma membrane might be regulated by PKC-down regulation without displacement to the cytosol and the interaction of p100 with the membrane might be reveersible.

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Study on the Antioxidative Activity of Human Milk (모유의 항산화능에 관한 연구)

  • 정해영;김정선;심경희;김명숙;김규원;이기영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.5
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    • pp.651-657
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    • 1995
  • The antioxidative activity of each fraction in human milk was examined using H2O2 and FeSO4-induced lipid peroxidation of mouse liver homogenate in order to elucidate the antioxidative substances of human milk. High molecular weight(~>20KD) fraction had more antioxidative effect on lipid peroxidation than low molecular weight(~ <20KD) fraction. Furthermore, the changes of antioxidative enzyme activities were estimated during lactation to study the roles of human milk. The human milk showed high activities of catalase, glutathione(GSH) peroxidase and GSH S-transferase. These results suggest that the antioxidative activities may mostly be attributed to high molecular weight fraction containing catalase, GSH peroxidase and GSH S-transferase.

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Soluble fraction from silk mat induced bone morphogenic protein in RAW264.7 cells

  • Kim, Seong-Gon;Jo, You-Young;Kweon, HaeYong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.41 no.2
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    • pp.51-55
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    • 2020
  • The objective of this study was to evaluate the changes in gene expression after incubation of cells with soluble fraction from different silk mat layers. A silk cocoon from Bombyx mori was separated into 4 layers of equal thickness. The layers were numbered from 1 to 4 (from the inner to outer layer). Each silk mat was placed into normal saline and collected soluble fraction. They were administered to RAW264.7 cells, and changes in the expression of genes were evaluated by cDNA microarray analysis. Layer 1 and 4 groups showed significantly higher expression of BMP-2 at 8 h after administration of soluble fraction (P < 0.05). Runx2 expression was significantly higher in Layer 4 group at 8h (P < 0.05). The silk mat from the innermost and outermost portion of the silkworm cocoon showed a significant change in the expression of genes that are associated with osteoinduction such as BMP-2 and runx2.

Effect of Ohmic Heating on Characteristics of Heating Denaturation of Soybean Protein (옴 가열이 대두 단백질의 열변성에 미치는 영향)

  • Cha, Yun-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.24 no.4
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    • pp.740-745
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    • 2011
  • Ohmic heating uses electric resistance heat which occurs equally and rapidly inside food when an electrical current is passed throught. In this study, we observed the physical & chemical characteristics changes which occurs in soybean protein during heating denaturation by using ohmic and conventional heating. After the ohmic heating process, we could not find any change of the primary protein structure in the denaturated soy protein samples. However, the rate of imbibed water(RIW) of the ohmic samples was 2 times faster than that of the conventional samples. Also the ANS-surface hydrophobicity was decreased, which is very closely related to RIW. In the differential scanning calorimeter(DSC) analysis result, all 7S soyprotein fraction samples were completely denaturated by ohmic and conventional heating. However, the 11S samples were completely denatured only by ohmic heating. According to the DSC result, we decided that soyprotein was damaged by temperature and electrical current during ohmic heating. The damage of electrical current was a cause of the characteristic changes.

STUDIES ON PROTEIN DEGRADABILITIES OF FEEDSTUFFS IN BANGLADESH

  • Khandaker, Z.H.;Tareque, A.M.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.6
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    • pp.637-642
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    • 1996
  • This experiment was conducted to determine RDP values of locally available feedstuffs that are commonly used in ruminant rations in Bangladesh. Four cattle were fistulated in the rumen for the in situ nylon bag studies. Seventeen different feedstuff sample (9 roughages and 8 concentrates) were evaluated in $4{\times}14cm$ nylon bags and incubated in the rumen for different periods of time (2, 6, 12, 24, 48 and 72 h). The variation in crude protein (CP) contents reflected on the average CP disappearance value throughout the rumen incubation. Soluble fraction (a), insoluble but degradable fraction (b) along with the rate of degradation also varied widely among the various feedstuffs. Under 2% of rumen outflow rate, the percentages of the calculated protein degradabilities of roughages were rice straw, 16.7; maize grass, 70.6; oat grass, 70.8; dhal grass, 71.1; sunhemp, 78.4; napier grass, 62.4; matikalai grass, 72.1; khesarikalai grass, 76.9 and daincha browse, 78.4, respectively. The results in the protein degradabilities (%) in 8% ruminal outflow rate of concentrates were wheat bran, 61.6; rice polish (red), 61.3; rice polish (auto), 30.9; mustard oil cake, 71.8; sesame oil cake, 74.2; coconut oil cake, 57.9; soybean meal, 49.2 and fish meal, 37.9, respectively.

Effects of Red Ginseng on Methyl Mercury Toxicities to Lipoprotein and Tissue Protein in Mouse (생쥐의 지단백질과 조직단백질에 미치는 메틸수은 독성에 대한 홍삼의 영향)

  • Chung, Hee Won;Soo Kyoung Shin;Choon Koo Lee
    • The Korean Journal of Ecology
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    • v.10 no.4
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    • pp.205-213
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    • 1987
  • In order to investigate the effects of red ginseng extract to methyl mercury toxicities in mice, the serum lipoproteins, tissure protein patternsm and growth rates were studied. Animals wee divided into 3 groups of the control, group I treated with methyl mercury chloride only, and group II treated together with methyl mercury chloride and red ginseng extract. In serum lipoprotein fractions of group I, beta lipoprotein fraction was increased and pre-beta lipoprotein fraction was decreased in comparision to those of the control. However, there was almost no difference in quantities of serum lipoprotein fractions between the control and group II. Total pretein contents of groups I and II were increased in liver and those of groups I and II in the kidney were decreased. However, in comparison to group I, total protein contents of group II in the liver and kidney were similar values with those of the control. Percentage of tissue protein fractions between control and group I in the liver and kindey showed considerable difference. On the other hand, the percentage of protein fractions of group II approximated to that of the control. Daily average growth rate of body weight in group II was similar to the control, but that of group I was decreased significantly in comparison to the other 2 groups.

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Biochemical Changes of Protein during the Senescence of Tobacco Leaf (담배잎의 노화과정에 따른 단백질의 생화학적 변화)

  • 이상각;심상인;강병화
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.5
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    • pp.563-568
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    • 1996
  • This experiment was conducted to obtain basic information of biochemical changes in the process of senescence by measuring the total RNA, protein, protease activity and electrophoretic pattern of protein in tobacco plant. The content of soluble protein increased by 15 days after leaf emergence and its level was not changed from 15 to 35 days after leaf emergence. The content of total RNA showed a maximum value at 15 days after leaf emergence and then decreased rapidly until 30 days after leaf emergence. The activity of protease of neutral fraction was higher than that of acidic fraction and rapidly increased up to the end of senescence after 50 days after leaf emergence. According to the analysis of electrophoresis, polypeptide band of 61kd was developed after 35 days after leaf emergence and increased by the end of senescence.

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Evaluation of Serum and Urine Protein Electrophoresis in Patients with Renal Disease (신장질환 환자에서 혈청과 요단백 전기영동 분획 평가)

  • Lim, Hyun-Jin;Back, Seong-Ok;Lee, Bum-Hee;You, Seon-Woo
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.2
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    • pp.111-116
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    • 2006
  • One of the cardinal findings of the renal diseases is proteinuria, which appears in the early phase of kidney diseases and is very important in diagnosis, prognosis and decision making in the treatment process and results of the treatment. The study subjects were 126 patients who visited the nephrology department of Kangbuk Samsung Hospital. Serum was requested for urine protein electrophoresis. Total protein was measured with Bayer Advia 1650 (Biuret). Quantitation of each fraction was done by multiplying the percentage of each fraction by the total protein. Serum creatinine and BUN were also measured with Bayer Advia 1650 (Jaffe and Urease). Serum protein EP was done with REP(rapid electrophoresis) using Helena Kit reagents (REP Ultra SPE Kit, Ponceau S stain, Acetic acid, Methanol, EP Control). Concentrated urine was used for urine protein EP. The SPSS package was used for statistics analysis. Percentage and quantitation of the level of albumin in renal diseases were significantly lower than those in healthy controls. Total protein was correlated with albumin. In terms of proportion, ${\alpha}1$-globulin, ${\alpha}2$-globulin, ${\beta}$-globulin, and ${\gamma}$-globulin fractions were increased in the disease group. But, in the quantified level, ${\alpha}2$-globulin was increased and ${\beta}$-globulin and ${\gamma}$-globulin were decreased. ESRD patients showed an increased secretion of high molecular proteins in urine protein EP. A decreased level in serum total protein correlated with the decreased level of serum albumin and the total amount of urine total protein. This study revealed the variety in the level of serum and urine proteins and their subgroups by EP.

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In vitro Comparision of Biological Activities of Solvent Fraction Extracts from Orostachys japonicus (와송(Orostachys japonicus) 용매별 분획 추출물의 항산화, 항균 및 암세포 독성 비교)

  • Kim, Seung Mi;Park, Jeong Hun;Boo, Hee Ock;Song, Sang Gi;Park, Hyeon Yong
    • Korean Journal of Plant Resources
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    • v.30 no.2
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    • pp.133-143
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    • 2017
  • This study was conducted to evaluate the contents of total polyphenol and flavonoid, and the effect of antioxidant, antimicrobial activities and cytotoxicity in vitro by different solvent fractions from Orostachys japonicus. The ethylacetate fraction extract for O. japonicus contained $634.48{\mu}g/g$ polyphenol and $205.20{\mu}g/g$ flavonoid. The ABTS radical scavenging ability of ethylacetate fraction extract at 1 mg/ml was higher than 95% which is comparable to ascorbic acid of 97%. The APX enzymatic activity and CAT activity were $1125.89{\mu}mol$ ascorbate oxidized/min/mg protein and 119.87 H2O2 decomposed/min/mg protein, respectively. In disc agar plate diffusion assay, the extract gave rise to a larger inhibition circle with Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus and Malassezia furfur strains compared with antibiotics kanamycin suggestive of high antibiotic activity. The cytotoxicity of extracts of O. japonicus was significant differences between solvent fractions. That is, the cytotoxic effect against human cancer cell was higher in ethylacetate fraction extract than other fraction extracts. These results suggest that fraction extract of O. japonicus might be very effective and economical in developing natural antioxidant and antimicrobial.

Detection of Protein Kinase C Isoenzymes in the Growth of Human Epidermal Keratinocytes by Growth Factors (Growth Factor를 처리한 피부상피세포로부터 Protein Kinase C Isoenzyme의 검출)

  • Eun-Young Joo;Nam-Woo Kim
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.83-91
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    • 2000
  • Subconfluent neonatal human epidermal keratinocytes were treated with a concentration 200 ng/$m\ell$ of human recombinant epidermal growth factor (hrEGF), human recombinant insulin-like growth factor-1 (hrIGF-1), and with a combination of hrEGF and hrIGF-1. Cytoplasmic and membrane-associated proteins were extracted and assayed. Proteins were separated by SDS-PAGE, and subjected to the western blot analysis. In the cytoplasmic fraction, the PKC concentration of keratinocyte treated with hrIGF-1 was higher than the control group, but the concentration of control group was the highest than the others in the membrane fraction. In the cytoplasmic fraction, EGF stimulated PKC-$\beta$II, -$\delta$, -$\theta$, and also stimulated PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$ and -$\theta$ in the membrane fraction. IGF-1 stimulated PKC-$\beta$I, -$\Im$ and -$\theta$ in the cytoplasmic, PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$, - $\varepsilon$ and -$\theta$ in the membrane. In the cells treated with a combination of EGF and IGF-1, PKC-$\alpha$, -$\beta$I, -$\Im$ and -$\theta$ in the cytoplasmic fraction, PKC-$\alpha$, -$\delta$, -$\Im$, -$\varepsilon$ and -$\theta$ in the membrane fraction were stimulated.

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