• Title/Summary/Keyword: Protein synthesis

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Continuous Cell-Free Protein Synthesis Using Glycolytic Intermediates as Energy Sources

  • Kim, Ho-Cheol;Kim, Tae-Wan;Park, Chang-Gil;Oh, In-Seok;Park, Kyung-Moon;Kim, Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.885-888
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    • 2008
  • In this work, we demonstrate that glycolytic intermediates can serve as efficient energy sources to regenerate ATP during continuous-exchange cell-free (CECF) protein synthesis reactions. Through the use of an optimal energy source, approximately 10 mg/ml of protein was generated from a CECF protein synthesis reaction at greatly reduced reagent costs. Compared with the conventional reactions utilizing phosphoenol pyruvate as an energy source, the described method yields 10-fold higher productivity per unit reagent cost, making the techniques of CECF protein synthesis a more realistic alternative for rapid protein production.

INFLUENCE OF PHENYLALANINE IN THE MEDIUM ON PROTEIN SYNTHESIS OF CHICKEN EMBRYO FIBROBLASTS

  • Kita, K.;Miyazaki, M.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.6
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    • pp.701-703
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    • 1996
  • The influence of phenylalanine (Phe) in the medium on protein synthesis of chicken embryo fibroblasts (CEF) was examined. CEF was derived from 9-d-old embryos by trypsin-EDTA digestion. To examine the deficiency of Phe in the medium, CEF was cultured in Dulbecco's modified Eagle's medium (DMEM) with or without Phe. CEF was also cultured in Dulbecco's phosphate buffered saline (PBS ($Ca^{2+}$, $Mg^{2+}$)) with or without $400{\mu}m$ Phe in order to examine the effect of Phe supplementation. All media were supplemented with 10% (v/v) fetal calf serum. After incubation for 6, 30 and 54 h, protein synthesis was measured by the incorporation of L-[2, $6-^{3}H$] Phe into CEF for further 18 h. Protein synthesis of CEF cultured in DMEM was higher than that in PBS ($Ca^{2+}$, $Mg^{2+}$). High specific radioactivity of Phe due to the low concentration of Phe in the medium resulted in the apparent increase in protein synthesis of CEF. Protein synthesis cultured in PBS ($Ca^{2+}$, $Mg^{2+}$) with Phe did not increase during 72 h of cell culture.

Inhibition of Corticosterone-induced Muscle Protein Synthesis by the Anabolic Steroid Nandrolone Phenylpropionate in Female Rats (아나보릭스테로이드인 Nandrolone Phenylpropionate가 암컷 쥐에서 코티코스테론에 의해 야기된 근육단백질 쇠퇴와 근육단백질 합성율 감소에 미치는 영향)

  • 주종재
    • Journal of Nutrition and Health
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    • v.29 no.8
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    • pp.867-873
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    • 1996
  • This study was undertaken to determine whether the anabolic steroid nandrolone phenylpropionate(NPP) can inhibit the muscle atrophy and reduction in muscle protein synthesis caused by glucocorticoids in female rates. Daily injections of 50mg/kg of corticosterone for eight days induced significant reductions in body weight gain and protein without affecting food intake. The mass, protein and RNA content, ratio of RNA to protein, and fractional rate of protein synthesis, measured in vivo, of gastrocnemius muscle were all significantly reduced by corticosterone treatement. Simultaneous administration of NPP at a dose of 10mg/kg with corticosteorne (50mg/kg) fully inhibited the reductions in the mass, protein and RNA content of gastrocnemius muscle, and body weight gain and protein with no alteration in food intake but the reduction in fractional rate of muscle protein syntheis was only partially prevented. The results indicate that the anabolic steroid nandrolone phenylpropionate is capable of preventing muscle atrophy in female rats treated with excess corticosterion.

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A Study of Mode of Action of Alachlor - I. Effects of Alachlor on Nucleic acid, Amino acid and Protein Synthesis in Oat(Avena sativa L.) (Alachlor의 제초기구(除草機構)에 관한 연구(硏究) - I.Alachlor가 귀리의 핵산(核酸), 아미노산 및 단백질합성(蛋白質合成)에 미치는 영향(影響))

  • Kwon, S.W.;Kim, J.C.
    • Korean Journal of Weed Science
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    • v.10 no.3
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    • pp.227-232
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    • 1990
  • The effects of alachlor [2-chloro-2', 6' diethyl-N-(methoxymethyl) acetanilide] treatment on nucleic acid, amino acid and protein synthesis were studied. The amide herbicide alachlor blocks the biosynthesis of the amino acids isoleucine, valine and aromatic amino acid in oat root tips. Nucleic acid was inhibited, but was not proportional to reduction in protein synthesis. $1{\times}10^{-4}M$ of alachlor treatment of oat roots inhibited 36% DNA synthesis, but DNA synthesis was not inhibited at $1{\times}10^{-5}M$. RNA synthesis was inhibited by $1{\times}10^{-5}M$ and $1{\times}10^{-4}M$ of alachlor 16 and 27%, respectively, while inhibition of protein synthesis did occur at same concentrations. Inhibition of protein synthesis also did not occur at concentration below $1{\times}10^{-4}M$ alachlor. It suggest that inhibition of protein sythesis caused significantly by alachlor($1{\times}l0^{-3}M$) result from secondary action.

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Influence of Refeeding with Vitamin, Mineral and Fibre on Protein Synthesis and Messenger Ribonucleic Acid Content in the Liver and Muscle of Fasted Chicks

  • Aman Yaman, M.;Kita, K.;Pinontoan, R.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.5
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    • pp.545-549
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    • 1998
  • The influence of refeeding with either vitamin, mineral, fibre of water on protein synthesis and mRNA content in the liver and breast muscle of fasted chicks was investigated. At 15 d of age, chicks were fasted for 2 d and then refed either vitamin, mineral, fibre or water. The fractional synthesis rate (FSR) of protein was measured after 30 min of refeeding by using a large dose injection of L - 2, $6[^3H]$ phenylalanine. In the liver, FSR was reduced by fasting and tended to increase but not significantly by refeeding with vitamin or mineral. FSR was not affected by refeeding with fibre or water. There was no influence of fasting and refeeding on ribosomal capacity (the RNA : protein ratio) and ribosomal efficiency (total protein synthesised per total RNA). The absolute synthesis rate (ASR) of liver protein and hepatic mRNA content were reduced by fasting and unchanged by refeeding. In the muscle, FSR, ASR and mRNA content were significantly decreased by fasting and not recovered by refeeding with either vitamin, mineral, fibre or water. It concluded that vitamin, mineral, fibre and water have little capacity to stimulate liver and muscle protein synthesis reduced by fasting.

Effects of the Protein Fraction of Panax ginseng on Primary Cultured Chicken Skeletal Muscle Cells (인삼 단백분획물이 일차배양한 계배의 근육세포에 미치는 영향)

  • Park, Mi-Jung;Song, Jin-Ho;Lee, Heun-Pa;Kim, Young-Choong
    • Korean Journal of Pharmacognosy
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    • v.21 no.3
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    • pp.210-216
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    • 1990
  • Effects of the protein fraction of Panax ginseng on chicken embryonic skeletal muscle cells cultured with a decfiient medium were studied. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton(fraction A), between 5,000 and 10,000 dalton(fraction B), between 1,000 and 5,000 dalton(fraction C), between 500 and 1,000 dalton(fraction D). According to the microscopic observation, all four protein fractions at the concentration of $10{\sim}100{\;}{\mu}g/ml$ showed the tendency to stimulate the growth and differentiation of the muscle cells. The activity of acetylcholinesterase in muscle cells was significantly elevated by the protein fraction A at the concentration of $100{\mu}{\;}g/ml$. Protein fractions B,C and D significantly enhanced the synthesis of RNA in the muscle cells. The synthesis of DNA in muscle cells was significantly enhanced by protein fractions A,B and C.

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Development and Application of the Paper Model of Protein Synthesis Process in High School Biology (단백질 합성 과정 종이 모형 개발과 고등학교 생물 수업에서의 활용)

  • Byun, Sun-Young;Shim, Kew-Cheol
    • Journal of Science Education
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    • v.34 no.2
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    • pp.268-278
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    • 2010
  • The purpose of this study was to develop the paper model of protein synthesis process in order for students to learn more effectively the protein synthesis, and to examine learning effects of instruction using it in high school biology. For this study, 117 students of 12th grade were sampled from a high school in Daejeon metropolitan city. The students were divided into two groups; the control group(n=58) were taught the protein synthesis by explanation-based traditional teaching method, and the experimental group(=59) were taught them by using protein synthesis model. Instruction using the model of protein synthesis process was more effective in improving the students' academic achievements and motivating interests in their learning than the traditional intruction. The class using the models can be another effective teaching method to teach students abstract and complicated concepts like protein synthesis process.

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Effects of Heavy Metals on Growth and Protein Synthesis in Cyanobacterium synechocystis sp. PCC 6803 (중금속이 Cyanobacterium synechocystis sp.PCC 6803의 성장과 단백질 합성에 미치는 영향)

  • 강경미;장남기
    • Asian Journal of Turfgrass Science
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    • v.10 no.4
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    • pp.315-329
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    • 1996
  • The changes of growth and protein synthesis pattern by aluminum (Al), cadmium (Cd), zinc (Zn) treatments were studied in Cyanobacterium synechocystis sp. PCC 6803. When exposed to Al from 5ppm to 3oppm, synechocystis grows normally. But more than that retard the growth of algae notably. The 0.05ppm Cd additions had no effect on the growth of algae. 0.1, 0.2, and 0.5ppm Cd inhibited growth. Under 1 and 2ppm Cd stress, growth was greatly diminished. Zn had dual effects. The growth of algae in media containing 5ppm was stimulated. As concentration increases more than l5ppm, growth inbition increases. Under 25ppm Zn stress, growth was greatly diminished. According to logistic theory, r and K values of each heavy metal-treated groups were estimated. Correlation analysis of r and K values with metal concentration shows that there is negative correlation between K and concentration in Cd and Zn treatments. Critical concentration which shows lethal or sublethal effect was estimated by t-test of each r and K value. The cells cultured in 10, 20, 30, 40 and 5oppm of Al, 1 and 2ppm of Cd, and 10, 15, 20, 25 and 30ppm of Zn for 4 days was used for protein analysis. Analysis of protein synthesis with SDS-PACE showed alterations of protein synthesis pattern. The synthesis of protein about 220kD increased markedly. In this study, it showed that resistance mechanism against Al, Cd, and Zn is K selection and that metal stress induced the change of protein synthesis in Cyanobacterium synechocystis sp. PCC 6803.Key words:Cyanobacterium synechocystis sp. FCC 6803, Heavy metals, Aluminum, Cadmiutm Zinc, Crowth, Frotein synthesis.

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THE EFFECT OF NATURAL EXTRACTS ON CELL GROWTH AND CYTOKINE PRODUCTION (생약 추출물이 세포성장 및 cytokine 생산에 미치는 영향)

  • Ryu, In-Cheol;Son, Seong-Heui;Chung, Chong-Pyoung;Bae, Ki-Hwan
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.37-47
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    • 1993
  • The native connective tissue attachment of the periodontium is known to be a complex consisting of gingival fibroblasts, periodontal ligament cells, gingival epithelial cells, cementum, alveolar bone and extensive extracellular matrix (collagen, glycoprotein and proteoglycans). The purpose of this study was to evaluate the effects of natural extracts on DNA, collagen and protein synthesis and inhibition of cytokine production in the gingival and periodontal ligament fibroblasts and gingival epithelial cells. Healthy gingival tissue was obtained from orthodontic treatment patients, and gingival epithelial cells, gingival fibroblasts and periodontal ligament cells were isolated and cultured from the samples. After treated with Ginseng protein, Pluronic F-68, Scutellariae Radix, centella asiatica, PDGF, IGF, DNA synthesis, total protein and collagen synthesis, and cytokine production of gingival epithelial cell, gingival fibroblast and periodontal ligamentcells were measured. MTT method for DNA synthesis, Peterkofsky and Dingerman method for total protein and collagen synthesis, and IL-1 ELISA kit for cytokine production were used. The proliferation of epithelial cells was enhanced in Centella asiatica, Ginseng protein, Pluronic F-68 and Scutellariae Radix. The activities of PDL cells were increased in PDGF, IGF, and Pluronic F-68. Higher collagen synthesis was observed in Scutellariae Radix and total protein synthesis was increased in Scutellariae Radix and PDGF. The inhibitory effects on IL-1, IL-6, $TNF-{\alpha}$ were observed in all exrracts.

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Effect of Protein Sources on Rumen Microbial Protein Synthesis Using Rumen Simulated Continuous Culture System

  • Joo, J.W.;Bae, G.S.;Min, W.K.;Choi, H.S.;Maeng, W.J.;Chung, Y.H.;Chang, M.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.3
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    • pp.326-331
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    • 2005
  • A rumen simulated continuous culture (RSCC) system was used to study the influence of supplementation of the three different types of protein sources such as urea, casein and soy protein on rumen microbial synthesis in terms of rumen microbial synchronization. The urea treatment showed the highest pH value. Ammonia nitrogen concentration was rapidly increased after feeding and not significantly different in the urea treatment (13.53 mg/100 ml). Protozoa numbers were not significantly different for soy protein and casein treatment compared to urea treatments during incubation. The average concentration of total VFA (mMol) was not detected with significant difference among treatments, but iso-butyrate production showed the highest for soy protein treatment among treatments (p<0.001). The lowest concentration in total iso-acids (iso-butyrate and iso-valerate) production was observed in urea treatment. The soy protein treatment showed no significantly change in acetate/propionate. The amounts of dry matter (DM) out flow showed no significant difference among treatments. Organic matter (OM) flow was the highest for urea treatments and the lowest for casein treatment (p<0.03). The nitrogen flow for casein treatment was not significantly different from other treatments. The efficiency of microbial protein synthesis in terms of microbial nitrogen (MN) synthesis (g MN/kg ADOM) digested in the rumen was highest for casein treatment (58.53 g MN/kg ADOM) compared to soy protein and urea (p<0.05). This result suggests that rumen ammonia releasing rate may influence on microbial protein synthesis in the rumen.