• Title/Summary/Keyword: ATPase6

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Direct Evidence of Intracellular Alkalinization in Saccharomyces cerevisiae KNU5377 Exposed to Inorganic Sulfuric Acid

  • Yun, Hae-Sun;Paik, Sang-Kyoo;Kim, Il-Sup;Jin, Ing-Nyol;Sohn, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.243-249
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    • 2004
  • The toxicity of inorganic sulfuric acid as a stressor was characterized in Saccharomyces cerevisiae KNU5377. In this work, we examined physiological responses to low extracellular pH $(pH_{ex})$ caused by inorganic $H_2SO_4$, which could not affect cell growth after pH was adjusted to an optimum with Trizma base. The major toxicity of sulfuric and was found to be reduction of environmental pH, resulting in stimulation of plasma membrane ${H^+}-ATPase$, which in turn contributed to intracellular alkalinization. Using a pH-dependent fluorescence probe, 5-(and-6)-carboxy SNARF-1, acetoxymethyl ester, acetate (carboxy SNARF-1 AM acetate), to determine $pH_{in}$, we found that color was dependent on the changes of intracellular pH which coincided with calculated $pH_{in}$ of alkalinization up to approximately pH 7.3. This alkalinization did not seem to affect survival of these cells exposed to 30 mM sulfuric acid, which lowered the $pH_{ex}$ of the glucose containing growth media up to approximately pH 3.0; however, the cells could grow only up to 70% of the maximum growth in the same media, when 30 mM sulfuric acid was added.

새로운 프로톤 펌프 억제제, IY-81233의 항위염과 항궤양작용 (Antigastric and Antiulcerative Action of a New Proton Pump Inhibitor (IY-81233))

  • 김승희;김진;강석연;이송득;홍성걸;김동연;문애리
    • Biomolecules & Therapeutics
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    • 제4권3호
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    • pp.285-290
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    • 1996
  • This study was designed to determine the effect of newly synthesized antiulcer agent, 5-pyrrolyl-6-halo-2-(pyridyl-2-methylthio)benzimidazole derivatives (IY-81233), on various experimental ulcers and on the secretion of prostaglandin $E_2(PGE_2)$ into the gastric lumen of rat. IY-81233 was previously reported to have a strong inhibitory effect on $H^+/K^$-ATPase and on gastric acid secretion in rats. Oral administration of IY-81233 at concentrations of 0.2, 2.0, and 20 mg/kg inhibited gastric lesions and duodenal ulcer induced by indomethacin, HCI-ethanol, water-immersion stress, cysteamine, and acetic acid in a dose dependent manner. Their IC$IC_{50}$ values were 3.4, 1.4, 0.8, 1.3, and 1.2 mg/kg, respectively. These results indicate that IY-81233 is a potent antiulcer agent although it is slightly less potent than omeprazole in healing of gastritis and ulcers. The secretion of $PGE_2$ into gastric lumen was also investigated in relation to the cytoprotective effect by IY-81233 in rats. The $PGE_2$ level was not changed significantly by an oral administration of IY-81233, suggesting that IY-81233 has little effect on the gastric protection. Therefore, it can be concluded that IY-81233 exerts prominent antiulcer activity by suppressing gastric acid secretion via an inhibition of a proton pump and not by protecting the gastrointestinal mucosa against various ulcerative stimuli.

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Response of broiler chickens to diets containing different levels of sodium with or without microbial phytase supplementation

  • Akter, Marjina;Graham, Hadden;Iji, Paul Ade
    • Journal of Animal Science and Technology
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    • 제61권2호
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    • pp.87-97
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    • 2019
  • Phytate induced excessive mineral excretion through poultry litter leads to poor performance and environmental pollution. Exogenous microbial phytase supplementation to poultry diets reduce the environmental excretion of nutrient and improve bird's performance. However, excessive dietary sodium (Na) level may hinder the phytase-mediated phytate hydrolysis and negate the beneficial effects of phytase. Therefore, this experiment was conducted to investigate the effects of different concentration dietary Na on phytase activity and subsequent impact on broiler performance, bone mineralisation and nutrient utilisation. In this study, six experimental diets, consisting of three different levels of Na (1.5, 2.5, or 3.5 g/kg) and two levels of microbial phytase (0 or 500 U/kg) were formulated by using $3{\times}2$ factorial design. The six experimental diets were offered to 360 day-old Ross 306 male chicks for 35 days, where, each experimental diet consisted of 6 replicates groups with 10 birds. Along with growth performance, nutrient utilization, intestinal enzyme activity, dry matter (DM) content of litter and mineral status in bone were analysed. Dietary Na and phytase had no effect on bode weight gain and feed intake. Birds on the low Na diet showed higher (p < 0.05) feed conversion ratio (FCR) than the mid-Na diets. High dietary Na adversely affected (p < 0.001) excreta DM content. Phytase supplementation to the high-Na diet increased (p < 0.01) the litter ammonia content. High dietary Na with phytase supplementation improved ($Na{\times}phytase$, p < 0.05) the AME value and ileal digestibility of Ca and Mg. The total tract retention of Ca, P, and Mg was reduced with high Na diet, which was counteracted by phytase supplementation ($Na{\times}phytase$, p < 0.001). The diets containing mid-level of Na improved (p < 0.001) the function of Na-K-ATPase and Mg-ATPase in the jejunum. The overall results indicate that high dietary Na did not affect phytase activity but influenced the nutrient utilization of birds, which was not reflected in bird overall performance.

흰쥐에서 인도메타신으로 유발된 위염에 대한 까마귀쪽나무열매추출물의 보호효과 (Protective effect of Litsea japonica fruit flesh extract on indomethacin-induced gastritis in rats)

  • 박성환;박인재;윤지현;최구희;김현정;서윤희;조주현
    • 한국식품저장유통학회지
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    • 제24권7호
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    • pp.1017-1024
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    • 2017
  • 본 연구에서는 까마귀쪽나무열매추출물(LJF-HE)의 흰쥐모델에서 인도메타신으로 유발되어지는 위염에 대한 억제효과를 확인하고자 하였다. 까마귀쪽나무열매추출물 투여그룹(LJF-HE-L, LJF-HE-M, LJF-HE-H)에서 염증길이와 위액량이 control 그룹에 비하여 통계적으로 유의하게 감소한 결과를 얻었다. 또한 위액량의 유의적인 감소효과로 위산분비가 억제되어 공격인자 작용이 약해지는 원인과 펩신의 활성도를 낮추는 원인으로 인하여 위염발생을 억제하는 것으로 판단되어진다. 까마귀쪽나무열매추출물(LJF-HE)의 위산 분비 억제 기능은 gastrin 및 histamine에 의해 발현되는 CCK-2r와 H2r 유전자의 발현을 억제하여, gastrin 및 histamine에 의한 위산 생성 단계를 조절하여, proton pump인 H+/K+ ATPase 유전자 발현을 억제시키며, 그 결과로 인하여 위산 분비를 억제하는 것으로 판단되어진다. 그리고 까마귀쪽나무열매추출물은 점액을 증가시켜 위 점막을 보호하는 PGE2의 함량을 높여, 위 점막 보호 기능을 나타내고 있으며, 더불어 염증성 cytokine인 TNF-${\alpha}$와 IL-$1{\beta}$의 생성을 낮춰주어 염증 매개반응을 저해하는 것으로 판단되어진다. 이와 같은 결과를 종합하면 까마귀쪽나무열매추출물(LJF-HE)이 인도메타신으로 유발되어지는 위염에 대한 억제효과가 있는 것으로 판단되었다.

Subtraction 기법을 이용한 한우 성장 단계 특이 발현 유전자 탐색 (Identification of the Differentially Expressed Genes of Hanwoo During the Growth Stage by Subtractive cDNA Hybridization)

  • 장요순;김태헌;윤두학;박응우;정일정;조진기
    • Journal of Animal Science and Technology
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    • 제44권1호
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    • pp.13-22
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    • 2002
  • 한우의 성장단계 특이발현 유전자를 탐색하기 위하여, 본 연구에서는 유전자의 발현 유무 및 발현정도의 차이를 나타내는 유전자를 분리하는데 있어 가장 강력한 수단으로 알려진 subtractive cDNA hybridization 기법을 이용하여 한우 등심조직으로부터 12개월령 및 24개월령 특이적인 subtractive cDNA library를 제작하였다. 성장단계 특이적인 유전자를 탐색하기 위하여, 6, 12 및 24개월령 cDNA를 사용하여 reverse northern blot 분석을 실시하였으며, 6개월령 cDNA probe에 대하여 특이적인 signal을 나타낸 3개의 clone은 EPV 20, Ca2+ ATPase, 및 TCTP 유전자와 유사성을 나타내었다. 12개월령 cDNA probe에 대하여 특이적인 signal을 나타낸 9개의 cDNA clone은 각각 VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein p1, ADP/ATP translocase T1 및 UCP 2 유전자와 높은 homology를 가지고 있었다. 또한 2개의 clone이 각각 12개월령 및 24개월령 cDNA probe에 대하여 특이적인 signal을 나타내었는데, 12개월령 cDNA probe에 대해서만 signal을 나타낸 clone은 ferrochelatase 유전자와 유사하였으며, 24개월령 probe에 대해서만 signal을 나타낸 clone은 ADRP 유전자와 유사하였다. 이상에서와 같이, 본 연구에서 제작한 성장단계 특이적인 subtractive cDNA library를 분석하여 14종의 유전자를 한우 성장단계 특이 발현 후보 유전자로 선정하여 염기서열을 분석하였으며, 이외에도 성장단계에 있어 특이적으로 발현될 것으로 추정되는 cDNA 클론의 염기서열을 분석하였다.

방어 Myosin과 갈색띠 매물고둥 Paramyosin의 물리화학적 성질 (Physicochemical Characteristics of Yellowtail Myosin and Whelk Paramyosin)

  • 변재형;최영준
    • 한국식품과학회지
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    • 제19권1호
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    • pp.29-37
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    • 1987
  • 수산동물의 근육단백질 중 특히 많은 분포를 보이며. 생리적으로 뿐만 아니라 식품학적으로도 중요한 myosin과 paramyosin을 방어 보통육과 갈색띠 매물고둥에서 추출하여 각각의 물리 화학적 성질을 비교 검토 하였다. 방어 보통육 myosin은 분자량 $4.6{\times}10^5\;dalton$, 278 nm 에서의 분자흡광계수 $B^{1cm}_{1%}$ 5.44, 고유점도 1.60dl/g 및 그 소수성에 따른 형광강도는 740이었으며, 용해도에 비추어 등전점은 pH 5.0부근이었다. 한편 갈색띠 매물고둥 paramyosin은 분자량 $2{\times}10^5\;dalton$, 278nm 에서의 분자흡광계수$(B^{1cm}_{1%})$ 3.04, 고유점도 2.60dl/g 및 그 소수성에 의한 형광강도는 468이었으며, 등전점은 pH5.6부근이었다. 그리고 방어 myosin의 ${Ca}^{2+}-ATPase$ 활성은 0.342 ${\mu}moles-Pi/min/mg-protein$이었고, 분자당 40개의 SH기를 함유하고 있었다. 아미노산 조성의 분석 결과, 방어 보통육 myosin과 갈색띠 매물고둥 paramyosin의 총아미노산의 잔기수는 단백질 1 mole당 각각 4030잔기와 1694잔기이었으며, 극성 대 비극성 아미노산의 비율은 0.54와 0.47이었고, 산성 대 염기성 아미노산의 비율은 1.64와 2.42이었다.

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Expression, Purification and Characterization of the BLM binding region of human Fanconi Anemia Group J Protein

  • Yeom, Kyuho;Park, Chin-Ju
    • 한국자기공명학회논문지
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    • 제20권1호
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    • pp.22-26
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    • 2016
  • FANCJ is a DNA helicase which contributes genome stability by resolving G-quadruplex DNA from 5' to 3' direction. In addition to main ATPase helicase core, FANCJ has the protein binding region at its C-terminal part. BRCA1 and BLM are the binding partner of FANCJ and these protein-protein interactions contribute genomic stability and the proper response to replication stress. As the first attempt for studying FANCJ-BLM interaction, we prepared BLM binding region of FANCJ and characterized with CD and NMR spectroscopy. FANCJ (881-941) with N-ter 6xHis was purified as the oligomer. Secondary structure prediction based on CD data revealed that FANCJ (881-941) composed with ${\beta}$ sheet, turn and coils.$^1H-^{15}N$ HSQC spectra showed nonhomogeneous peak intensities with less number of peaks comparing than the number of amino acids in the construct. It indicated that optimization should be necessary for detailed further structural studies.

Multiple Age-Associated Mitochondrial DNA Deletions in Mouse Brain

  • Kim, Jin-Sun;Kim, Min-Jung;Kwon, In-Sook;Song, Eun-Sook
    • BMB Reports
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    • 제30권1호
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    • pp.33-36
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    • 1997
  • Age-dependent deletion of mitochondrial DNA (mtDNA) was detected in mouse brain using PCR method. The size of the deleted fragment was 0.5 kb, 0.9 kb. 1.7 kb and 4.3 kb in the region between cytochrome b gene and ATPase 6 gene. The deleted fragment was increased gradually from 3-month to 22month Direct repeat sequence flanking the deletion in 0.5 kb PCR product was TAAT.

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Expression of Latent P-Type ATPases and Their Presumptive Roles in Cell Membrane of Helicobacter pylori

  • YUN, SOON-KYU;SE-YOUNG HWANG
    • Journal of Microbiology and Biotechnology
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    • 제7권6호
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    • pp.378-385
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    • 1997
  • Cation motive ATPases on cell membranes of Helicobacter pylori were investigated using everted membrane vesicles. Latent ATPases could be ascertained from aggregated vesicle using N, N-dimethylformamide (DMF) and Triton X-100. By contrast, ultrasonication or chloroform treatments caused membranes to be disrupted, resulting in an alteration of sensitivities against azide or vanadate. Considerable amounts of vanadate-sensitive enzymes were identified from vesicle micelles, prepared by the dilution method. These were activated in the presence of either $Ni^{2+}\;or\;NH_4^+$. From studies employing H. pylori intact cell systems, we found that ATPase expression of this bacterium was markedly dependent upon air composition. It was interesting that cellular expression of $Ni^{2+}$- or $NH_4^{+}$-motive ATPases was significantly affected by extracellular pH, suggesting that these unique enzymes may physiologically be involved in cellular $Ni^2$ import and $NH_4^+$ export, respectively.

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유선조직내에 출현하는 dendritic cell의 형태학적 연구 II. 전자현미경적 관찰 (Morphological studies on the dendritic cells in the mammary gland II. Electron microscopic observations)

  • 류시윤;이차수
    • 대한수의학회지
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    • 제28권2호
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    • pp.241-249
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    • 1988
  • In order to investigate the morphological characteristics of dendritic cells in the mammary gland of the mouse (C57 BL/6), rat(W), rabbit and cat, the fine structures of the dendritic cells have been observed by the electron microscope. The results obtained were summarized as follows: The dendritic cells with the well-developed processes had an irregular shape, and lacked the desmosome. The pinocytotic vesicles and tubular invaginations of the cell membrane were frequently observed, and the mitochondria with the well-developed cristae were located in the restricted region in the cytoplasm of the dendritic cells. The nuclei of dendritic cells were indented, In the mice and rats, the dendritic cells had a few Langerhans cell granules(Birbeck granules). From the above results, it is confirmed that the ATPase-positive dendritic cells in the mammary gland are the Langerhans cells.

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