• Title/Summary/Keyword: cytosol

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Abscisic Acid Binding to Extracts from Normal and Viviparous-1 Mutant Aleurone Layers of Zea mays L.

  • Bai, Dong-Gyu
    • Journal of Plant Biology
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    • v.37 no.2
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    • pp.151-158
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    • 1994
  • Aleurone layers of normal and vp1 mutant maize kernels were extracted and centrifuged at 100,000g to yield a cytosol fraction. Binding of [3H]ABA cis, trans (+)ABA to a soluble macromolecular components present in the cytosol was demonstrated by Sephadex chromatography and non-denaturing PAGE. The binding component was of high molecular weight and seems to be an aggregate of proteins. A rapid DEAE-cellulose filter method for assaying bound [3H]ABA to a soluble protein was adapted. Binding assays were performed with cytosol that had been preheated or incubated with several enzymes, indicating that heat and protease treatments disrupted the binding. This suggested that binding occurred to proteins. Some properties of the ABA binding proteins were described. The [3H]ABA binding were reduced dramatically when unlabeled ABA was added as a competitor, suggesting a specific binding of [3H]ABA. Gel filtration profiles and autoradiogram of [3H]ABA binding showed no difference in the binding components of Vp1 and vp1/vp1 mutant cytosol, indicating that Vp1 protein is not a sole ABA binding protein.

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Studies on the Properties of lntracelluar Arsenic Binding Substances in the Rat Liver (흰쥐 간조직의 세포내 비소결합물질의 특성에 관한 연구)

  • 최임순;부문종;김충현
    • The Korean Journal of Zoology
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    • v.33 no.4
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    • pp.476-492
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    • 1990
  • Sodium arsenite (As) aqueous solution containing 4 ppm wss given to male rats for 15 days as drinking water. Electrophoretic pattern of liver cytosol from As-fed rats appeared to be significantly different from that of the control rats. Although the normal protein content of the cytosol fraction of As-fed rat liver was decreased, 8 stress proteins were increased. In liver cytosol fractions of As-treated rat, one kind of arsenic-binding substance (AsBS) was observed. Molecular weight of AsBS was identified to be 500 D and composition of amino acid was glycine, glutamic acid and cysteine. Glutathione (GSH) appeared to bind to arsenic and GSH-As complex showed the same mobility as AsBS on gel filtration chromatography. GSH conjugated As prevented As from inhibiting respiration, conformational change and swelling-contration of mitochondria. According to the above results. it is concluded that in vfuo treated arsenic stimulated synthesis of stress protein, and arsenic-binding substance might be glutathione and have a protective role against arsenic toxicity.

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Subcellular Distribution of Heavy Metals in Organs of Bivalve Modiolus Modiolus Living Along a Metal Contamination Gradient

  • Podgurskaya, Olga V.;Kavun, Victor Ya.
    • Ocean Science Journal
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    • v.41 no.1
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    • pp.43-51
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    • 2006
  • Concentration and distribution of Fe, Zn, Cu, Cd, Mn, Pb, Ni among subcellular fractions (cellular membrane structures and cytosol) and Zn, Cu, Cd among cytoplasmic proteins in the kidney and digestive gland of mussel Modiolus modiolus living along a polymetallic concentration gradient were studied. It was found in the kidney of M. modiolus from contaminated sites that the Fe percent increased in the "membrane" fraction, whereas Zn, Pb, Ni and Mn percent increased in the cytosol compared to the kidney of the control mussel. Note kidney cytosol of M. modiolus from clean and contaminated sites sequestered major parts of Cu and Cd. In the digestive gland of M. modiolus from contaminated sites Fe, Zn, Cd, Mn, Ni percent increased in the "membrane" fraction, whereas Cu, Pb percent increased in the cytosol compared to digestive gland of control mussel. Gel-filtration chromatography shows kidney of M. modiolus contains increased metallothionein-like protein levels irrespective of ambient dissolved metal concentrations. It was shown that the metal detoxification system in the kidney and digestive gland of M. modiolus was efficient under extremely high ambient metal levels. However, under complex environmental contamination in the kidney of M. modiolus, the metal detoxification capacity of metallothionein-like proteins was damaged.

A Study on Antiestrogenic Effects of Tamoxifen in Immature Rat Uterus; I. Effects on Concentrations of Cytosol and Nuclear Estradiol Receptor (미성숙 쥐 자궁에서 Tamoxifen의 Antiestrogen 효과에 관한 연구 : I. 세포질 내 및 핵 내 Estradiol 수용체 농도의 변화에 관하여)

  • Lee, Hyo-jong;Jo, Choong-ho;Park, Moo-hyun
    • Korean Journal of Veterinary Research
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    • v.25 no.2
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    • pp.187-195
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    • 1985
  • The Present study has been carried out to elucidate the antiestrogenic effects of tamoxifen in uteri of immature rats. Immature female Sprague-Dawley rats were allocated into 4, groups and injected with $5{\mu}g$ of estradiol-$17{\beta}$, $50{\mu}g$ of tamoxifen, a combination of both or vehicle only subcutaneously three times after an interval of 24 hours respectively. The concentrations, of cytosol estradiol receptor in uterus were measured by DCC method before and 1, 3, 6, 12, 24, 48 and 72 hours after the above treatments and those of nuclear estradiol were measured by protamine exchange method 72 hours and those of nuclear estradiol were measured by protamine exchange method 72 hours after the above treatments. The results obtained were summarized as follows: 1. The binding affinity of tamoxifen to estradiol receptor in uterine cytosol was lower than that of estradiol-$17{\beta}$, accordingly the translocation of estradiol receptor into the nucleus was found to be delayed. 2. Tamoxifen caused the retention of estradiol receptor in nucleus over 24 hours and inhibited the replenishment of the receptor from nucleus to cytosol in uterus.

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N-terminal formylmethionine as a novel initiator and N-degron of eukaryotic proteins

  • Kim, Jeong-Mok
    • BMB Reports
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    • v.52 no.3
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    • pp.163-164
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    • 2019
  • The ribosomal synthesis of proteins in the eukaryotic cytosol has always been thought to start from the unformylated N-terminal (Nt) methionine (Met). In contrast, in virtually all nascent proteins in bacteria and eukaryotic organelles, such as mitochondria and chloroplasts, Nt-formyl-methionine (fMet) is the first building block of ribosomal synthesis. Through extensive approaches, including mass spectrometric analyses of the N-termini of proteins and molecular genetic techniques with an affinity-purified antibody for Nt-formylation, we investigated whether Nt-formylated proteins could also be produced and have their own metabolic fate in the cytosol of a eukaryote, such as yeast Saccharomyces cerevisiae. We discovered that Nt-formylated proteins could be generated in the cytosol by yeast mitochondrial formyltransferase (Fmt1). These Nt-formylated proteins were massively upregulated in the stationary phase or upon starvation for specific amino acids and were crucial for the adaptation to specific stresses. The stress-activated kinase Gcn2 was strictly required for the upregulation of Nt-formylated proteins by regulating the activity of Fmt1 and its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins could be distinct N-terminal degradation signals, termed fMet/N-degrons, and that Psh1 E3 ubiquitin ligase mediated the selective destruction of Nt-formylated proteins as the recognition component of a novel eukaryotic fMet/N-end rule pathway, termed fMet/N-recognin.

Subcellular Localization of Catalase Encoded by the ctl+ Gene in Schizosaccharomyces pombe

  • Lee, Sang-il;Lee, Joon;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.156-159
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    • 2000
  • The cttl+ gene in Schizosaccharomyces pombe encoeds a catalse responsible for H2O2-resistance of this organism as judged by the H2O2-sensitive phenotype of the ctt1Δ mutant. In this study, we investigated the subcellular localization of the Ctt1 gene product. In wild type cells catalase activity was detected in the organelle fraction as well as in the cytosol. The ctt1Δ mutant contained no catalase activity, indicating that both cytosolic and organellar catalases are the products of a single ctt1+ gene. Western bolt analysis revealed two catalase bands, both of which disappeared in the ctt1Δ mutant. The major, fastermigrating band existed in the cytosol whereas the monor, slower-migrating band appeared to be located in organelles, most likely in peroxisomes. These results suggest that the ctt1+ gene product targeted to the peroxisome is a modified form of the one in the cytosol.

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Effect of Silkworm Powder on Oxygen radicals and Their Scavenger Enzymes in Brain membranes of SD Rats (뇌조직의 활성산소 및 그 제거효소에 미치는 누에분말의 영향)

  • 최진호;김대익;박수현;김동우;김정민;이희삼;류강선
    • Journal of Sericultural and Entomological Science
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    • v.42 no.2
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    • pp.93-98
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    • 2000
  • This study was designed to investigate the effect of silkworm powder on oxygen radicals and their scavenger enzymes in brain membrances of SD rats. Hydroxyl radical (OH) levels resulted in a considerable decreases in brain mitochondria fraction. Superoxide radical (O$_2$) levels were a slightly decreased in brain cytosol fraction. Lipid peroxide (LPO) and Oxidized protein (OP) levels were significantly decreased in brain mitochondria and microsomes fraction. Mn-superoxide dismutase (SOD) activity was remarkably increased in the mitochondria fraction. Cu and Zn-SOD activities were effectively increased in brain cytosol fraction. GSHPx activity was considerably increased in brain cytosol fraction. These results suggest that anti-aging effect of silkworm plays an effective role in attenuating an oxidative stress and increasing a scravenger enzyme activity in brain membranes.

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A Study on the Glucose-regulating Enzymes and Antioxidant Activities of Water Extracts from Medicinal Herbs (한약재의 물 추출물이 당대사 관련 효소와 항산화 활성에 관한 연구)

  • Choe, Myeon;Kim, Dae-Jung;Lee, Hyeon-Ju;You, Jin-Kyoun;Seo, Dong-Joo;Lee, Joon-Hee;Chung, Mi-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.5
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    • pp.542-547
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    • 2008
  • The anti-diabetic effects of water extracts (WE) from medicinal herbs on hepatic glucose-regulating enzymes, such as glucokinase (GCK), pyruvate dehydrogenase (PDH), acetyl-CoA carboxylase (ACC) and ${\alpha}$-glucosidase, were studied using the cytosol fraction in liver and mitochondia fraction in heart of a type II diabetic animal (GK rat, Goto-Kakizaki). The free radical scavenging activity of water extracts by DPPH method was also tested. We found that free radical scavenging activity was strong in Corni fructu (CF), Mokdan Bark (MDB), Chenhwabon (CHB) and Sanyack (SY), while that of Backbocreng (BBR), Shuckgihwang (SGH) and Taecsa (TS) was lower. For GCK activity in cytosol of liver, CF and CHB had a more effective activity than other extracts. PDH activity in mitochondria fraction of heart was higher in all of the extracts, expect for the TS extract, than in the control. ACC activity in cytosol fraction of liver was significantly higher in the CF, CHB, SGH, TS and SY extracts than in the control. CF, BBR and MDB led to a decrease in the ${\alpha}$-glucosidase activity. Therefore, these results suggest that all of the extracts may be used as functional material in the development as anti-diabetic functional food and medicine.

Protective effects of Gagaminjakdowha-Tang on liver injury of rats (흰쥐의 간손상(肝損傷)에 대한 가감인작도화탕(加減茵芍桃花湯)의 간(肝) 보호효과)

  • Kang Jae-Chun;Kim Byong-Woo;Lee Tae-Houn
    • The Journal of Internal Korean Medicine
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    • v.24 no.1
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    • pp.44-54
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    • 2003
  • Objectives : This study was done to investigate the protective effects of Gagaminjakdowha-Tang on liver injury of rats induced by $CCl_4$ and d-galactosamine. Methods : All animals were divided into 5 groups, those were normal group(untreated), control group(treated with 0.9% Saline solution), sample I group(200mg/kg administrated), sample II group(400mg/kg administrated), Silymarin(200mg/kg administrated) group. Liver injury of rats were induced by $CCl_4$ and d-galactosamine, and then the serumtransaminase(ALT & AST) alkaline phosphatase(ALP), lactic dehydrogenase(LDH) for enzyme activities, Liver cytosol malondialdehyde(MDA), catalase, superoxide dismutase(SOD), glutathione S-transferase(GST) and glutathione-peroxidase(GPX) for enzyme activities were measured. Results : The inhibitory effects on the serum ALT activities were noted in both sample I and sample II group. The inhibitory effects on the serum AST activities were noted in only sample II group. The inhibitory effects on the serum ALP activities were noted in both sample I and sample II group. The inhibitory effects on the serum LDH activities were noted in only sample II group. The inhibitory effects on the liver cytosol malondialdehyde were noted in only sample II group. The decresed effects on the liver cytosol catalase activities were inhibited in only sample II group. The decresed effects on the liver cytosol superoxide dismutase activities were inhibited in only sample II group. The decresed effects on the liver cytosol GST activities were inhibited in only sample II group. The decresed effects on the liver cytosol GPX activities were inhibited in only sample II group. The inhibitory effects of the serum ALT activities were noted in both sample I and sample II. The inhibitory effects of the serum AST activities were noted in only sample II group. The inhibitory effects of the serum ALP activities were noted in only sample II group. The inhibitory effects of the serum LDH activities were noted in both sample I and sample II group. Conclusions : Gagaminjakdowha-Tang has protective effects against liver injury in rats induced by $CCl_4$ and d-galactosamine.

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

  • 최진호;김대익;박수현;김정민;백영호;이희삼;류강성
    • Journal of Life Science
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    • v.10 no.5
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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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