• Title/Summary/Keyword: $H^+-ATPase$

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The Study of Trnascriptional Regulated Gene, $hrp^{2+}$, in Yeast

  • Choi, In-Soon
    • Journal of Life Science
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    • v.11 no.2
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    • pp.111-115
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    • 2001
  • This study was designed to clone the SNF2/SW12 helicase-related genes from the fission yeast Schizosaccha-romyces pombe and thereafter to elucidate the common functions of the proteins in this family. The $hrp^{2+}$gene was cloned by polymerase chain reaction amplification using degenerative primers from conserved SNF2 motifs within the ERCC6 gene, which encodes a protein involved in DNA excision repair. Like other SNF2/SW12 family proteins, the deduced amino acid sequence of Hrp2 contains DNA-dependent ATPase/7 helicase domains as well as the chromodomain and the DNA binding domain. This configuration is similar to that of mCHD1 (mouse chromo-ATPase/helicase-DNA-dinding protein 1), suggesting that Hrp2 is a S. pombe homolog of mCHD1, which is thought to function in altering the chromatin structure to control the gene expression. To characterize the function of Hrp2, 4 Uracil-Hrp2 fusion protein, it was purified near homogeneity by affinity chromatography on $Ni^{2+}$-NTA agarose, DEAE-Sepharose ion exchange arid Sephacryl S-200 gel filtration chromatographies. The purified fusion protein exhibited DNA-dependent ATPase activity, which was stimulated by both double-stranded and single-stranded DNA. To determine the steady-state level of $hrp^{2+}$ transcripts during growth, cells were cultured in medium and collected at every 2hr to prepare total RNAs. The northern blot analysis showed that the level of $hrp^{2+}$ transcripts reached its maximum before the cells entered the exponential growth phase and then decreased gradually, This result implies that Hrp2 may be required at early stages of cell growth.h.

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$[^3H]$ Ouabain Binding and Effect of Ouabain on $^{45}Ca^{2+}$-Uptake in Rat Cardiac Myocytes (쥐 심근 세포의 $[^3H]$ Ouabain 결합과 $^{45}Ca^{2+}}$섭취에 미치는 Ouabain의 영향)

  • 이신웅;김영희;진갑덕
    • YAKHAK HOEJI
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    • v.28 no.3
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    • pp.129-138
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    • 1984
  • Specific [$^{3}H$] ouabain binding and $Ca^{2+}$ -uptake were measured to elucidate the role of high affinity [$^{3}H$] ouabain binding site in rat cardiac myocytes which contain 65% of rod cells. High affinity [$^{3}$H] ouabain binding site, which is about 3% of total pump sites, with apparent dissociation constant ($K_{D}$) of $1.1{\times}10^{-7}M$ and maximum binding site concentration (Bmax) of 1.2 pmol/mg protein ($1.754{\times}10^{5}cells$) were identified. At the concentration of $10^{-7}M$ to $10^{-4}M$, ouabain produced concentration dependent increase in $Ca^{2+}$-uptake of myocytes. The effect of ouabain on $Ca^{2+}$-uptake was not effected by membrane depolarization (elevated K+ in incubation medium) or verapamil. These results suggest that in rat ventricular myocytes the ouabain receptor complex to high affinity site may increase Na+ - $Ca^{2+}$ exchange across the sarcolemmal membrane by inhibition of Na+, K+ - ATPase.

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Genetic improvement of potato plants

  • Suharsono, Sony
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.12-12
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    • 2017
  • Genetic improvement in potato can be carried out through several approaches, as sexual crosses, somatic hybridization, mutation and genetic engineering. Although the approach is different, but the goal is the same, to get a superior cultivar. Mutation and genetic engineering are very interesting methods for genetic improvement of potato plants. Mutation by gamma-ray irradiation have been performed to get some new potato cultivars which are more resistant to disease and have higher productivity. We have carried out a mutation of some potato cultivars and obtained some excellent clones to be potentially released as new superior cultivars. By the mutation method, we have released one potato cultivar for the French fries industry, and we registered one cultivar of potato for chips, and two cultivar for vegetable potatoes. Actually we are doing multi-location trial for three clones to be released as new cultivars. Through genetic engineering, several genes have been introduced into the potato plant, and we obtained several clones of transgenic potato plants. Transgenic potato plants containing FBPase gene encoding for fructose bisphosphatase, have a higher rate of photosynthesis and higher tuber productivity than non-transgenic plants. This result suggests that FBPase plays an important role in increasing the rate of photosynthesis and potato tuber productivity. Some transgenic potatoes containing the Hd3a gene are currently being evaluated for their productivity. Over expression of the Hd3a gene is expected to increase tuber productivity and induce flowering in potatoes. Transgenic potato plants containing MmPMA gene encoding for plasma membrane ATPse are more tolerant to low pH than non-transgenic plants, indicating that plasma membrane ATPase plays an important role in the potato plant tolerance to low pH stress. Transgenic potato plants containing c-lysozyme genes, are highly tolerant of bacterial wilt diseases caused by Ralstonia solanacearum and bacterial soft rot disease caused by Pectobacterium carotovorum. Expression of c-lyzozyme gene plays an important role in increasing the resistance of potato plants to bacterial diseases.

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Quality Characteristics of Mackerel Surimi Prepared by Alkaline Washing under Reduced Pressure (감압 알칼리 수세하여 제조한 고등어 Surimi의 품질 특성)

  • Park, Hyung-Sun;Park, Sang-Woo;Yang, Seung-Taek
    • Korean Journal of Food Science and Technology
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    • v.30 no.5
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    • pp.1120-1127
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    • 1998
  • An attempt was made in this study to investigate the optimum condition of washing for preparation of mackerel surimi by alkaline washing of 1, 3, 5, and 7 times under atmospheric (760), 660, and 560 mmHg pressure. The qualities of surimis were examined by analyzing the factors such as water content, crude lipid, pH, volatile basic nitrogen (VBN), expressible drip, protein extractability, $Mg^{2+}-$, $Ca^{2+}-$ and EDTA-ATPase activity, transglutaminase (TGase) activity, gel strength and color. The contents of moisture, crude lipid, pH and VBN in surimis prepared by alkaline washing under atmospheric, and reduced pressure went up to $72.0{\sim}72.9%$, $4.8{\sim}5.7%$, $6.9{\sim}7.0$ and $6.7{\sim}7.0\;mg/100\;g$, respectively. Protein extractability, ATPase activity and TGase activity were highest in surimis prepared by alkaline washing under 560 mmHg. Gel strengths of surimi setting gel and cooked gel from five times washing under 560 mmHg were 420 g cm (atmospheric, 330 g cm) and 485 g cm (atmospheric, 412 g cm), respectively. For the preparation of mackerel surimi, optimum washing condition was five times washing under 560 mmHg.

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The Effect of Salviae Radix on Oxidat-Inhibition of Phosphate Uptake in Renal Proximal Tubular Cells (단삼약침액(丹蔘藥鍼液)이 신장(腎臟) 근위세뇨관세포(近位細尿管細胞)에서 산화제(酸化劑)에 의한 인산(燐酸)의 이동억제(移動抑制)에 미치는 영향(影響))

  • Lee, Ho-Dong;Youn, Hyoun-Min;Jang, Kyung-Jeon;Song, Choon-Ho;Ahn, Chang-Beohm
    • Journal of Acupuncture Research
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    • v.17 no.3
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    • pp.208-218
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    • 2000
  • This study was undertaken to determine if Salviae Radix (SR) exerts protective effect against oxidant-induced inhibition of phosphate uptake in renal proximal tubular cells. Membrane transport function and cell death were evaluated by measuring phosphate uptake and trypan blue exclusion, respectively, in opossum kidney (OK) cells, an established proximal tubular cell line. $H_2O_2$ was used as a model oxidant. $H_2O_2$ inhibited the phosphate uptake in a dose-dependent manner over the concentration range of 0.1-0.5 mM. Similar fashion was observed in cell death. However, the phosphate uptake was more vulnerable to $H_2O_2$ than cell death, suggesting that $H_2O_2$-induced inhibition of phosphate uptake is not totally attributed to cell death. Decreasedphosphate uptake was associated with ATP depletion and inhibition of $Na^+$-pump activity as determined by direct inhibition of $N^+-K^+$-ATPase activity. When cells were treated with $H_2O_2$ in the presence of 0.05% SR, the inhibition of phosphate uptake and cell death induced by $H_2O_2$ was significantly attenuated. SR restored ATP depletion and decreased $Na^+-K^+$-ATPase activity, and this is likely responsible for the protective effect of SR on decreased phosphate uptake. The protective effect of SR was similar to the $H_2O_2$ scavenger catalase. SR reacts directly with $H_2O_2$ to reduce the effective concentration of the oxidant. The iron chelator deferoxamine prevented the inhibition of phosphate uptake and cell death induced by $H_2O_2$, suggesting that $H_2O_2$-induced cell injury is resulted from an iron-dependent mechanism. These results indicate that SR exerts the protective effect against $H_2O_2$-induced inhibition of phosphate uptake by reacting directly with $H_2O_2$ like the $H_2O_2$scavenger enzyme catalase, in OK cells. However, the underlying mechanism remains to be explored.

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Ultrastructural and Cytochemical Studies on the Endometrial Stromal Cells of Guinea Pig during Estrous Cycle (발정주기(發情週期)에 따른 Guinea Pig의 자궁내막(子宮內膜) 기질세포(基質細胞)의 미세구조(微細構造)및 세포화학적(細胞化學的) 연구(硏究))

  • Deung, Young-K.;You, Kwan-H.;Choi, Choon-K.;Lee, Choon-K.;Chung, Ho-S.
    • Applied Microscopy
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    • v.12 no.1
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    • pp.57-68
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    • 1982
  • Cyclical changes in the fine structures of endometrial stroma of guinea pig during the estrous cycle were studied by electron microscopy. Cytochemical studies were made in order to investigate the ultrastructural localization of the acid phosphatase,alkaline phosphatase and ATPase in these cells. The results obtained are as follows: 1. During estrus collagen fibers were most abundant in the stroma. The stromal cells showed increases in the number of several cytoplasmic organelles, especially the rough endoplasmic reticulum was significantly increased and the structures were greatly differentiated. 2. Many cytoplasmic processes and cell debris have been distributed in the stroma of metestrus. The distributions were increased and degenerated mitochondria were observed during diestrus. 3. Cytochemical studies indicated that during metestrus and diestrus acid phosphatase activities were localized in the degenerating collagen fibers. Alkaline phosphatase activities were weak in the collegen fibers during proestrus and estrus which intense activities were localized around the cell membrane during metestrus and diestrus. ATPase activities were present on the cell membrane and intercellular space of stromal cell during proestrus and estrus.

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Inhibition of $N^{+}-K^{+}$ Adenosine Triphosphatase Activity in Fisher Rats by Uranyl Nitrate

  • Lee, Kee-Ho;Lee, Je-Ho;Lee, Soo-Yong;Park, Sang-Yoon;Lee, Seung-Hoon;Yun, Taik-Koo;Ryu, Young-Wun;Lim, In-Kyoung
    • Journal of Radiation Protection and Research
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    • v.15 no.2
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    • pp.1-6
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    • 1990
  • An attempt was made to test the possibility of a major role for the $Na^{+}-K^{+}$ adenosine triphosphatase (ATPase)system in the diuresis induced by uranyl nitrate(UN). Fisher 344 rats were intravenously injected with UN(5 mg/kg, 15 mg/kg and 30 mg/kg). Urinary excretion of $Na^{+}\;and\;K^{+}$ significantly increased in 24 h exposure on the UN and then decreased below the normal level 3 days after the treatment. $Na^{+}-K^{+}$ ATPase activity of kidney was significantly inhibited in high dosages of UN 15mg/kg and UN 30 mg/kg 3-5 days after injection. And then the recovery of the enzyme activity was observed within 5-10 days after injection, at which the regeneration of the tubular cells occurred.

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Up-Regulation of RANK Expression via ERK1/2 by Insulin Contributes to the Enhancement of Osteoclast Differentiation

  • Oh, Ju Hee;Lee, Na Kyung
    • Molecules and Cells
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    • v.40 no.5
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    • pp.371-377
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    • 2017
  • Despite the importance of the receptor activator of nuclear factor (NF)-kappaB ligand (RANKL)-RANK signaling mechanisms on osteoclast differentiation, little has been studied on how RANK expression is regulated or what regulates its expression during osteoclastogenesis. We show here that insulin signaling increases RANK expression, thus enhancing osteoclast differentiation by RANKL. Insulin stimulation induced RANK gene expression in time- and dose-dependent manners and insulin receptor shRNA completely abolished RANK expression induced by insulin in bone marrow-derived monocyte/macrophage cells (BMMs). Moreover, the addition of insulin in the presence of RANKL promoted RANK expression. The ability of insulin to regulate RANK expression depends on extracellular signal-regulated kinase 1/2 (ERK1/2) since only PD98059, an ERK1/2 inhibitor, specifically inhibited its expression by insulin. However, the RANK expression by RANKL was blocked by all three mitogen-activated protein (MAP) kinases inhibitors. The activation of RANK increased differentiation of BMMs into tartrate-resistant acid phosphatase-positive ($TRAP^+$) osteoclasts as well as the expression of dendritic cell-specific transmembrane protein (DC-STAMP) and d2 isoform of vacuolar ($H^+$) ATPase (v-ATPase) Vo domain (Atp6v0d2), genes critical for osteoclastic cell-cell fusion. Collectively, these results suggest that insulin induces RANK expression via ERK1/2, which contributes to the enhancement of osteoclast differentiation.

$Ca^{2+}$ Effect on Conversion of Exogenous 1-Aminocyclopropane-1-Carboxylic Acid to Ethylene in Vigna radiata Protoplasts

  • Seung-Eun Oh
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.271-276
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    • 1994
  • The possibility that 1-aminocyclopropane-1-carboxylic acid (ACC)-uptake may be dependent on the H+-gradient established across the plsma membrane was tested in protoplasts isolated from 2.5 day old mungbean hypocotyls. The ACC-induced ethylene production was inhibited when the H+-gradient was collapsed by the treatment with carbonycyamide-p-trifluro-methoxy-phenylhydrazone (FCCP). Moreover, the treatment with o-vanadate, a specific inhibitor of plasma membrane H+-ATPase, caused the inhibition of ethylene production. The ACC-induced ethylene production was inhibited by the treatemnt with verapamil (Ca2+-channel blocker), or ethylene glycol-bis($\beta$-aminoethyl ether) N, N, N', N'-tetraacetic acid (EGTA) (Ca2+-chelator). In contrast, the ehtylene production was stimulated by the application of A23187 (Ca2+ ionophore). The inhibitory effect of EGTA in the ethylene producton was magnified in the presence of A23187. From these results, we suggest that the external Ca2+ influx to the cytosol resulted in the stimulatin of ACC oxidase activity after ACC-uptake resulting from a H+-gradient across the plasma membrane.

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Reconstitution of Membrane Proteins from Corn Seedlings with Phosphatidylcholine (옥수수 유식물 조직에서 분리한 막 단백질과 Phosphatidylcholine의 재조합)

  • 오승은
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.321-323
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    • 1990
  • Membrane proteins isolated from the coleoptile and mesocotyl tissues of corn seedlings were solubilized with Triton X100 and reconstituted with phosphatidylcholine at 2$0^{\circ}C$. The proteoliposomes were incubated and proton uptake into the vesicles was measured with a spectrophotometer. Addition of ATP to the reaction mixture was found to result in an active accumulation of proton into the vesicles. These results indicate that the preparation contains tightly bound phosphatidylcholine vesicles with reconstituted H+ -ATPase from the plant cell membranes.

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