• Title/Summary/Keyword: total proteins

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A Comparative Study of Protein Profiles in Porcine Fetus Fibroblast Cells with Different Confluence States

  • Han, Rong-Xun;Kim, Hong-Rye;Diao, Yunfei;Kim, Myung-Youn;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.243-248
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    • 2009
  • To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

Food Consumption and Nutrition Survey in Korean Rural Areas (농촌(農村) 식품섭취실태(食品攝取實態) 및 영양조사(營養調査))

  • Park, Yaung-Ja;Chun, Sung-Kyu
    • Journal of Nutrition and Health
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    • v.9 no.2
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    • pp.87-97
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    • 1976
  • This study is based on data from the food consumption survey on 727 members of 125 farm households from 7 different provinces. The survey was conducted in May, 1975 in cooperation with the O.R.D. The results obtained in this study are summarized as follows. 1. The average consumption of the basic food groups per capita per day was 563 g for cereals and grains(398g of rice and 129g of barley), 87.6g for meats and legumes, 317.8g for fruits and vegetables, 25.7g for milks and small fishes, 9.1g for fats and oils, and 45.1g for other group. 2. The average daily consumption of calories and nutrients was 2256 cal and 11.7g for animal proteins, 70.5g for total proteins, 21.6g for fats, 537.4mg for calcium, 18.1mg for iron, 5375lU for vitamin A, 1.27mg for thiamine, 1.05mg for riboflavin, 15.5mg for niacin, 77.7mg for ascorbic acid. When these figures are compared with the recommended allowances for Korean, the calories and nutrients intakes were satisfactory, except for the intakes of animal protein which was below two third of the recommended allowance. 3. The diets of the projected villages differed from those of the non-projected villages in the following respect: (a) The amounts of animal proteins and fats were larger in the projected villages than in the non-projected villages. (b) The percentage contribution of fats to the total amount of calories from three nutrients, carbohydrates, proteins and fats was higher in the projected villages than in tile non-projected villages. (c) The percentage contribution from carbohydrates to the total amount of calories was higher in the non-projected villages than in the projected villages. 4. Certain physical and clinical symptoms were observed among the people in the rural areas, which can be related to the shortages of animal proteins and fats in their diets. It is recommended to pay special attention to the nutrition of school children in the Korean rural areas.

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Changes of haemolymph proteins in Pieris rapae L. during the cuticle formation and hardening process (배추흰나비의 큐티클 形成과 硬化에 따른 혈림프 단백질의 變化)

  • Hak Ryul Kim;Eul Won Seo
    • The Korean Journal of Zoology
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    • v.23 no.1
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    • pp.1-12
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    • 1980
  • Changes and possible origin of haemolymph proteins during the cuticle formation and hardening are determined by means of acrylamide gel electrophoresis and immunodiffusion. The results by acrylamide gel electrophoresis showed at least 19 protein bands in the haemolymph and 13 fractions in the fat body with relatively constant pattern during the period of cuticle formation and hardening. Both haemolymph and fat body proteins are generally characterized by the presence of three to four heavy stained bands and several thin bands near the top region of the gel. At least over five haemolymph proteins are constantly present during this period. Immunodiffusion tests show that of total eight to nine pupal haemolymph proteins two proteins were already detected in the fat body before pupation and other two proteins were also found in the fat body immediately after pupation, suggesting fat body as possible source of these two haemolymph proteins.

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Comprehensive Proteome Analysis of the Excretory/Secretory Proteins of Toxoplasma gondii

  • Lee, Won-Kyu;Ahn, Hye-Jin;Baek, Je-Hyun;Lee, Chong-Heon;Yu, Yeon Gyu;Nam, Ho-Woo
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.3071-3076
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    • 2014
  • Proteomic analyses of the excretory/secretory proteins from the RH strain of Toxoplasma gondii have been performed to understand their functions in the host-parasite interaction. A total of 34 proteins were identified from LC/MS/MS analysis and their abundance was estimated by spectral counting methods. Among them, 8 species of micronemal proteins (MICs), 2 species of rhoptry proteins (ROPs), and 6 species of dense granular proteins (GRAs) were confirmed. Besides these, 18 species of protein were newly identified, and their cellular functions were estimated from sequence analysis. The three most abundant of the 34 identified extractor/secretory proteins-GRA1, GRA7 and GRA2-were confirmed to be highly expressed in T. gondii using the spectral count method. This phenomenon is another demonstration of the importance of GRA proteins for the penetration and survival of T. gondii.

Global analysis of ginsenoside Rg1 protective effects in β-amyloid-treated neuronal cells

  • Shim, Ji Seon;Song, Min-Young;Yim, Sung-Vin;Lee, Seung-Eun;Park, Kang-Sik
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.566-571
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    • 2017
  • Background: A number of reports have described the protective effects of ginsenoside Rg1 (Rg1) in Alzheimer's disease (AD). However, the protective mechanisms of Rg1 in AD remain elusive. Methods: To investigate the potential mechanisms of Rg1 in ${\beta}$-amyloid peptide-treated SH-SY5Y cells, a comparative proteomic analysis was performed using stable isotope labeling with amino acids in cell culture combined with nano-LC-MS/MS. Results: We identified a total of 1,149 proteins in three independent experiments. Forty-nine proteins were significantly altered by Rg1 after exposure of the cells to ${\beta}$-amyloid peptides. The protein interaction network analysis showed that these altered proteins were clustered in ribosomal proteins, mitochondria, the actin cytoskeleton, and splicing proteins. Among these proteins, mitochondrial proteins containing HSD17B10, AARS2, TOMM40, VDAC1, COX5A, and NDUFA4 were associated with mitochondrial dysfunction in the pathogenesis of AD. Conclusion: Our results suggest that mitochondrial proteins may be related to the protective mechanisms of Rg1 in AD.

In vitro Ccovalent Binding of SC-42867, PGE2 Antagonist, to Rat Liver Microsomal Proteins

  • Lee, Kyung-Tae
    • Archives of Pharmacal Research
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    • v.18 no.6
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    • pp.381-384
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    • 1995
  • Covalent binding of the reactive metabolites of SC_42867 to microsomal proteins has been examined. In the absence of inhibitor of cytochrome oxydase (.alpha.-naphtyl-isothiocyanate) or a radical scavenger (3-terthiobuty-4-hydroxyanisol), up to 4.0% of total redioactivity used in the assay could irreversibly bind to proteins. In the presence of an inhibitor, the highest percentage of covalent binding observed is 0.7% a significant decrease of the metabolism of SC42876 was observed. These results suggest in a cytochrome P-450 dependent generation of SC_42867 metabolites significantly take part in the covalent binding process.

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Interactions between secreted GRA proteins and host cell proteins across the parasitophorous vacuolar membrane in the parasitism of Toxoplasma gondii

  • Ahn, Hye-Jin;Kim, Sehra;Kim, Hee-Eun;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.44 no.4 s.140
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    • pp.303-312
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    • 2006
  • Interactions between GRA proteins of dense granules in Toxoplasma gondii and host cell proteins were analyzed by yeast two-hybrid technique. The cMyc-GRA fusion proteins expressed from pGBKT7 plasmid in Y187 yeast were bound to host cell proteins from pGADT7-Rec-HeLa cDNA library transformed to AH109 yeast by mating method. By the selection procedures, a total of 939 colonies of the SD/-AHLT culture, 348 colonies of the $X-\alpha-gal$ positive and PCR, 157 colonies of the $X-\beta-gal$ assay were chosen for sequencing the cDNA and finally 90 colonies containing ORF were selected to analyze the interactions. GRA proteins interacted with a variety of host cell proteins such as enzymes, structural and functional proteins of organellar proteins of broad spectrum. Several specific bindings of each GRA protein to host proteins were discussed presumptively the role of GRA proteins after secreting into the parasitophorous vacuoles (PV) and the PV membrane in the parasitism of this parasite.

Fractionation of enzymatically methylated acid-insoluble proteins from thymus nuclei

  • Lee, Hyang-Woo;Kim, Sang-Duk;Paik, Woon-Ki
    • Archives of Pharmacal Research
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    • v.9 no.3
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    • pp.157-161
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    • 1986
  • Isolated calf thymus nuclei were in vitro methylated with S- adenosy-L-methyl-$^{14}C$ methionine, and the proteins were fractionated according to their solubilities. Histone fraction ($H_{2}SO_{4}$-soluble fraction) contained approximately 60% total radioactivity incorporated, while "residual protein" which was ($H_{2}SO_{4}$-insoluble contained the remaining radio-activity. The "residual protein" was further fractionated into various acidic proteins, which contained very littel of the radioactivity. However, the protein fraction eluted from DEAE-cellulose with 0.5 N NaOH contained the largest amount of radioactivity. This protein was found to be basic in nature by amino analysis.

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Proteomic Approach Analysis of Mammary Membrane Proteins Expression Profiles in Holstein Cows

  • Yang, Yong-xin;Cao, Sui-zhong;Zhang, Yong;Zhao, Xing-xu
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.885-892
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    • 2009
  • To investigate host defense mechanisms for protecting the mammary gland from mastitis infection, the membrane fraction of mammary tissues from Holstein cows was purified by differential velocity centrifugation, and then the sodium dodecyl sulfate-polyacrylamid gel electrophoresis (SDS-PAGE) separated proteins were identified by ion trap mass spectrometer equipped with a Surveyor high performance liquid chromatography (HPLC) system. A total of 183 proteins were identified. Bioinformatics software was applied to analyse physicochemical characteristics of the identified proteins and to predict biochemical function. These data may provide valuable information to investigate the mechanisms of mammary gland milk secretion and infectious disease, and enable a clear identification of proteins and potential protein targets for therapies.

Development of new food protein through chemical modification of rice bran proteins

  • Bae, Dongho;Jang, In Sook
    • Journal of Applied Biological Chemistry
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    • v.42 no.4
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    • pp.180-185
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    • 1999
  • Protein concentrate was produced and succinylated from rice bran to assess and improve its functional properties for the purpose of expanding the uses of rice bran proteins. The most effective solvent for the extraction of rice bran proteins was 20% aqueous ethanol at pH 9. The protein content of rice bran protein concentrate produced was 70.0% and the total protein yield was 64.3%. The extent of succinylation of free amino groups in the modified products was 72.8%. Though the modified protein products showed good functional properties including solubility, emulsion properties, and oil absorption capacity, it did not form gel. Succinylation improved solubility and emulsion and gelling properties. These improvements in functionality will enhance the value of rice bran proteins, thus enabling them to be more competitive with other food proteins.

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