• 제목/요약/키워드: Oxidized protein

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Tracing Metabolite Footsteps of Escherichia coli Along the Time Course of Recombinant Protein Expression by Two-Dimensional NMR Spectroscopy

  • Chae, Young Kee;Kim, Seol Hyun;Ellinger, James J.;Markley, John L.
    • Bulletin of the Korean Chemical Society
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    • 제33권12호
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    • pp.4041-4046
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    • 2012
  • The recombinant expression of proteins has been the method of choice to meet the demands from proteomics and structural genomics studies. Despite its successful production of many heterologous proteins, Escherichia coli failed to produce many other proteins in their native forms. This may be related to the fact that the stresses resulting from the overproduction interfere with cellular processes. To better understand the physiological change during the overproduction phase, we profiled the metabolites along the time course of the recombinant protein expression. We identified 32 metabolites collected from different time points in the protein production phase. The stress induced by protein production can be characterized by (A) the increased usage of aspartic acid, choline, glycerol, and N-acetyllysine; and (B) the accumulation of adenosine, alanine, oxidized glutathione, glycine, N-acetylputrescine, and uracil. We envision that this work can be used to create a strategy for the production of usable proteins in large quantities.

광합성 세균인 Rhodopseudomonas gelatinosa ATCC 17013에서 Cytochrome c-551의 정체 (Purification of Cytochrome c-551 from Photosynthetic Bacterium Rhodopseudomonas Gelatinosa ATCC 17013)

  • 강대길;최원기
    • 미생물학회지
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    • 제29권2호
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    • pp.92-96
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    • 1991
  • The soluble cytochrome c-551 of photosynthetic bacterium, Rhodopseudomonas gelatinosa ATCC 17013 was purified through a sequene of four step chromatography including CM-cellulose ion-exchange chromatography, DEAE-Sephacel chromatography, Sephacryl s-200 gel permeation chromatography, and HPLC (SP-5PW). The molecular weight of the purified cytochrome c-551 was 14, 600 Da, and this protein shows the absorption peak at 551 nm, 522 nm, and 417 nm as the reduced form, and at 412 nm as the oxidized form. The cytochrome c-551 seems to be a substrate for the terminal oxidase in the electron transport chain.

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A 43 kD Protein Isolated from the Herb Cajanus indicus L Attenuates Sodium Fluoride-induced Hepatic and Renal Disorders in Vivo

  • Manna, Prasenjit;Sinha, Mahua;Sil, Parames C.
    • BMB Reports
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    • 제40권3호
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    • pp.382-395
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    • 2007
  • The herb, Cajanus indicus L, is well known for its hepatoprotective action. A 43 kD protein has been isolated, purified and partially sequenced from the leaves of this herb. A number of in vivo and in vitro studies carried out in our laboratory suggest that this protein might be a major component responsible for the hepatoprotective action of the herb. Our successive studies have been designed to evaluate the potential efficacy of this protein in protecting the hepatic as well as renal tissues from the sodium fluoride (NaF) induced oxidative stress. The experimental groups of mice were exposed to NaF at a dose of 600 ppm through drinking water for one week. This exposure significantly altered the activities of the antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR) and the cellular metabolites such as reduced glutathione (GSH), oxidized glutathione (GSSG), total thiols, lipid peroxidation end products in liver and kidney compared to the normal mice. Intraperitoneal administration of the protein at a dose of 2 mg/kg body weight for seven days followed by NaF treatment (600 ppm for next seven days) normalized the activities of the hepato-renal antioxidant enzymes, the level of cellular metabolites and lipid peroxidation end products. Post treatment with the protein for four days showed that it could help recovering the damages after NaF administration. Time-course study suggests that the protein could stimulate the recovery of both the organs faster than natural process. Effects of a known antioxidant, vitamin E, and a non-relevant protein, bovine serum albumin (BSA) have been included in the study to validate the experimental data. Combining all, result suggests that NaF could induce severe oxidative stress both in the liver and kidney tissues in mice and the protein possessed the ability to attenuate that hepato-renal toxic effect of NaF probably via its antioxidant activity.

The Dry-aging and Heating Effects on Protein Characteristics of Beef Longissiumus Dorsi

  • Kim, Ji-Han;Lee, Ha-Jung;Shin, Dong-Min;Kim, Tae-Kyung;Kim, Young-Boong;Choi, Yun-Sang
    • 한국축산식품학회지
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    • 제38권5호
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    • pp.1101-1108
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    • 2018
  • The aim of this study was to investigate the effects of dry-aging (DA) and the cooking process on the myofibril protein functionalities and in vitro digestibility of proteins in beef loin. Six sirloins from beef were dry-aged for 28 d, and the control group (n=6) was analyzed 2 d postmortem for this study. Dimensional changes (reduction of thickness and surface shrinkage) after cooking were significantly greater in the control group than the DA group, whereas the shear force of the DA group was significantly lower than that of the control. Effect of cooking on aggregation, hydrophobicity, and in vitro digestibility were significantly higher in the DA group than in the control. After cooking, the protein in DA sirloins was more oxidized than in the control samples. According to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis result, the low molecular weight bands (below 17 kDa) increased in the DA group, finding that the protein characteristics of dry-aged beef was affected by cooking.

Comparative study on Hsp25 expression in Mongolian gerbil and mouse cerebellum

  • Lee, Heang-Yeon;Kim, Seong-Hwan;Lee, Jae-Bong;Shin, Chang-Ho
    • 한국동물위생학회지
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    • 제29권4호
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    • pp.469-482
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    • 2006
  • The term 'heat shock protein (Hsps)' was derived from the fact that these proteins were initially discovered to be induced by hyperthermic conditions. In response to a range of stressful stimuli, including hyperthermia, immobilization, UV radiation, amino acid analogues, arsenite, various chemicals, and drugs the mammalian brain demonstrates a rapid and intense induction of the heat shock protein. Moreover, Hsps were expressed on the various pathological conditions including trauma, focal or global ischemia, hypoxia, infarction, infections, starvation, and anoxia. Especially, Hsp25 has a protective activity, facilitated by the ability of the protein to decrease the intracellular levels of reactive oxygen species (ROS) as well as its chaperone activity, which favors the degradation of oxidized proteins. Recently, it has clearly demonstrated that Hsp25 is constitutively expressed in the adult mouse cerebellum by parasagittal bands of purkinje cells in three distinct regions, the central zone (lobule VI-VII) and nodular zone (lobule IX-X), and paraflocculus. The Mongolian gerbil has been introduced into stroke study model because of its unique brain vasculature. There are no significant connections between the basilarvertebral system and the carotid system. This anatomy feature renders the mongolian gerbil susceptible to forebrain ischemia-induced seizure. The present study is designed to examine the pattern of Hsp25 expression in the cerebellum of this animal in comparison with that in mouse.

간장의 산화적 스트레스 및 세포막 유동성에 미치는 누에분말의 영향 (Effects of Silkworm (Bombyx mori L.) Powder on Oxidative Stress and Membrane Fluidity in Liver of SD Rats)

  • 최진호;김대익;박수현;김정민;조원기;이희삼;류강선
    • 생명과학회지
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    • 제10권5호
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    • pp.496-503
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    • 2000
  • This study was designed to investigate the effects of silkworm (Bombyx mori L.) powder on oxidative stress and membrane fluidity in liver membranes of rats. Sprague-Dawley (SD) male rats (160±10 g) were fed basic diet (control group), and experimental diets (SWP-200 and SWP-400 groups) added 200 and 400 mg/kg BW/day for 6 weeks. A significant differences between liver mitochondria and microsomes of SWP-200 and SWP-400 groups could not be obtained. Membrane fluidities were dose-dependently increased (14.8% and 28.5%, 20.0% and 29.9%) in liver mitochondria and microsomes of SWP-200 and SWP-400 groups compared with control group. Basal oxygen radicals (BOR) in liver mitochondria and mocrosomes were significantly inhibited (15.2% and 21.7%, 12.6% and 18.6%, respectively) by SWP-200 and SWP-400 groups compared with control group. Induced oxygen radicals (IOR) in liver microsomes were significantly inhibited (15.5% and 16.1%, respectively) by SWP-200 and SWP-400 groups compared with control group, but IOR in liver mitochondria was significantly inhibited about 12.0% by SWP-400 group only compared with control group. Lipid peroxide (LPO) levels were significantly decreased (14.4% and 9.1%, respectively) in liver mitochondria and microsomes of SWP-400 group only compared with control group. Oxidized protein (OP) levels were remarkably decreased about 12.7% and 16.3% in liver microsomes only of SWP-200 and SWP-400 groups, but significant difference between liver motochondria could not obtained. These results suggest that administration of SWP may play an effective role in a attenuating a oxidative stress and increasing a membrane fluidity in liver membranes.

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간장조직의 활성산소 및 그 제거효소에 미치는 뽕잎 추출물의 영향 (Effects of Mulberry (Morus alba L.) Leaf Extract on Oxygen Radicals and Their Scavenger Enzymes in Liver of SD Rats)

  • 최진호;김대익;박수현;김정민;백영호;이희삼;류강성
    • 생명과학회지
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    • 제10권5호
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    • pp.504-510
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    • 2000
  • This study was designed to investigate the effects of mulberry (Morus alba L.) leaf extract (MLE) on oxygen radicals and their scavenger enzymes in liver membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10 g) were fed basic diet (control group), and experimental diets (MLE-100 and MLE-300 groups) added 100 and 300 mg/kg BW/day for 6 weeks. Hydroxyl radical (.OH) levels resulted in a significant decreases (15.2% and 18.1%, 5.6% and 8.0%, respectively) in liver mitochondria and microsomes could be not obtained. These are no significant differences in superoxide radical ($O_2$) levels of liver cytosol in MLE-100 and MLE-300 groups compared with control group. Lipid peroxide (LPO) levels were slightly decreased about 13.6% and 6.1% in liver mitochondria and microsomes of MLE-300 group compared with control group. Oxidized protein (OP) levels were remarkably decreased about 16.9% and 27.2% in liver microsomes only of MLE-100 and MLE-300 group compared with control group. Mn-SOD activities in liver mitochondria were remarkably increased (18.2% and 28.7%, respectively) in MLE-100 and MLE-300 groups, and Cu,Zn-SOD activities in liver cytosol were also significantly increased (11.3% and 20.2%, respectively) in MLE-100 and MLE-300 groups compared with control group. Mn-SOD activities in liver mitochondria were remarkably increased (18.2% and 28.7%, respectively) in MLE-100 and MLE-300 groups, and Cu,Zn-SOD activities in liver cytosol were also significantly increased (11.3% and 20.2%, respectively) in MLE-100 and MLE-300 groups compared with control group, but significant difference between GSHPx activities in liver cytosol could be not obtained. These results suggest that anti-aging effect of mulberry leaf extract (MLE) may play a pivotal role in attenuating a various age-related changes.

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뇌조직의 산화적 스트레스 및 세포막 유동성에 미치는 실크 피브로인의 영향 (Effects of Silk Fibron on Oxidative Stress and Membrane Fluidity in Brain of SD Rats)

  • 최진호;김대익;박수현;김정민;이종수;이광길;여주홍;이용우
    • 생명과학회지
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    • 제10권5호
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    • pp.511-518
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    • 2000
  • This study was designed to investigate the effects of silk fibroin powder (SFP : Mw 500) on oxidative stress and membrane fluidity in brain membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10 g) were fed basic diet (control group), and experimental diets (SFP-2.5 and SFP-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. Cholesterol level was significantly decreased about 8.0% in brain microsomes of SFP-5.0 group only compared with control group. Membrane fluidities were significantly increased (12.9% and 15.2%, respectively) in brain microsomes of SFP-2.5 and SFP-5.0 groups, but significant difference between in brain mitochondria of these two groups could be not obtained. Basal oxygen radicals (BOR) in brain mitochondria and microsomes were significantly ingibited (10.4%, and 24.0%, 7.9% and 14.9%, respectively) by SFP-2.5 and SFP-5.0 groups compared with control group. Induced oxygen radicals (IOR) in brain mitochondria and microsomes were significantly inhibited (11.8% and 14.1%, respectively) by SFP-5.0 groups compared with control group compared with control group. Lipid peroxide (LPO) levels were dose-dependently decreased (12.9% and 21.9%, 13.2% and 22.5%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were significantly decreased (15.7% and 17.1%, 16.7% and 15.7%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-2.5 and SFP-5.0 groups compared with control group. These results suggest that administration of SFP may play an effective role in a attenuating a oxidative stress and increasing a membrane fluidity in brain membranes.

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동물실험을 통한 솔잎(松葉) 유효성분의 항노화효과 구명 및 구조 해명 I. 간장의 세포막 유동성과 산화적 스트레스에 미치는 솔잎 추출물의 영향 (Investigation of Anti-aging Effect and Determination of Chemical Structures of Pine Needle Extract (PNE) through the Animal Experiments I. Effects of PNE on Membrane Fluidity and Oxidative Stress in Liver of SD Rats)

  • 최진호;김대익;박수현;김동우;이종수;김현숙
    • 생명과학회지
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    • 제9권4호
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    • pp.473-480
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    • 1999
  • This study was designed to investigate the effects of pine (Pinus densiflora Sieb et Zucc) needle extract (PNE) on membrane fluidity and oxidative stress in liver membranes of Sprague-Dawley (SD) rats as a study on investigation of anti-aging effect and determination of chemical structures of PNE through the animal experiments. Male SD rats were fed basic diets (control group) and experimental diets (0.5% and 1.0%-PNE group) for 6 weeks. Administrations of 0.5% and 1.0%-PNE resulted in a marked decreases (15∼25% and 23∼26%, respectively) in cholesterol accumulations of liver mitochondria and microsomes compared with control group. Membrane fluidities were significantly increased (15∼25%) in liver microsomes of 0.5% and 1.0%-PNE groups compared with control group. Formations of basal and induced oxygen radicals (BOR and IOR) in liver mitochondria were significantly inhibited (11∼12% and 10∼15%, respectively) by administrations of 0.5% and 1.0%-PNE compared with control group. Lipid peroxide (LPO) levels were remarkbly decreased about 20% in liver mitochondria and microsomes of 0.5% and 1.0%-PNE groups compared with control group. Oxidized protein levels calculated with carbonyl group were significantly decreased about 15% in liver mitochondria of 1.0%-PNE group compared with control group. These results suggest that PNE may play a effective role in a attenuating a oxidative stress and increasing a membrane fluidity.

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Rat 간장의 산화적 스트레스 및 세포막 유동성에 미치는 실크 피브로인의 영향 (Effects of Silk Fibroin in Oxdative Stress and Membrane Fluidity in the Liver of SD Rats)

  • 최진호;김대익;박수현;김동우;이광길;여주홍;김정민;이용우
    • 한국잠사곤충학회지
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    • 제42권1호
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    • pp.58-64
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    • 2000
  • This study was designed to investigate the effects of silk fibroin powder (Mw 500) on oxidative stress and membrane fluidity in liver membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10g) were fed basic diet (control group), and experimental diets (SEP-2.5 and SFP-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. Cholesterol levels resulted in a significant decrease (12.1% and 9.0%, respectively) in the liver mitochondria and microsomes of SEP-5.0 group compared with control group. Membrane fluidity as significantly increased (16.1% and 16.5%, 5.8% and 17.4%) in the liver mitochondria and microsomes were significantly inhibited (16.1% and 18.3%, 8.1% and 15.1%, respectively) at the SFP-2.5 and SEP-5.0 groups compared with control group. Induced oxygen radicals (BOR) in liver mitochondria and microsomes were significantly inhibited (16.1% and 18.3%, 8.1% and 15.1%, respectively) at the SFP-2.5 and SEP-5.0 groups compared with control group. Induced oxygen radicals (IOR) in liver microsomes were significantly inhibited (17.0% and 26.6%, respectively) at the SFP-2.5 and SFP-5.0 groups compared with control group, but IOR in liver mitochondria was significantly inhibited about 12.3% at the SWP-400 group only compared with control group. Lipid peroxide (LPO) levels were significantly decreased (8.3% and 18.0%, 13.4% and 18.4%, respectively) in the liver mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were dose-dependently decreased (5.4% and 11.6%, 19.0% and 24.4%, respaectively) in the iver mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. These results suggest that administration of SFP may play an effective role in attenuating an oxidative stress and increasing a membrane fluidity in liver membranes.

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