• Title/Summary/Keyword: $H^+/K^$-ATPase

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Gastroprotective Activities of Sennoside A and Sennoside B via the Up-Regulation of Prostaglandin E2 and the Inhibition of H+/K+-ATPase

  • Hwang, In Young;Jeong, Choon Sik
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.458-464
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    • 2015
  • Sennoside A (erythro) and sennoside B (threo) are dianthrone glycosides and diastereomers. We investigated their abilities to prevent the gastric lesions associated with diseases, such as, gastritis and gastric ulcer. To elucidate their gastroprotective effects, the inhibitions of $HCl{\cdot}tOH$-induced gastritis and indomethacin-induced gastric ulcers were assessed in rats. It was observed that both sennoside A and sennoside B increased prostaglandin $E_2$ ($PGE_2$) levels and inhibited $H^+/K^+$-ATPase (proton pump). In a rat model, both compounds reduced gastric juice, total acidity and increased pH, indicating that proton pump inhibition reduces gastric acid secretion. Furthermore, sennoside A and B increased $PGE_2$ in a concentration-dependent manner. In a gastric emptying and intestinal transporting rate experiment, both sennoside A and sennoside B accelerated motility. Our results thus suggest that sennoside A and sennoside B possess significant gastroprotective activities and they might be useful for the treatment of gastric disease.

Purification and Characterization of a 15S Arpase from Chick Skeletal Muscle (계 골격근에서 15S ATPase의 순수분리 및 특성연구)

  • 심규석;채광수
    • The Korean Journal of Zoology
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    • v.37 no.4
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    • pp.524-530
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    • 1994
  • An Arpase complex has been purified to apparent homogeniety from the extract of chick skeletal muscle using conventional column chromatographies and glycerol density gradient centrifugation. This eWe has a sedimentation coefficient of 15S as determined by the gradient centrifugation and therefore is referred to as the 15S ATPase. It behaves as a 600-kOa molecule upon gel filtration analysis using a Superose-6 column. However, the ATPase runs as a 95-kDa polvpeptide when analyzed by polvacrvlamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, the Arpase is likely to consist of six identical subunits of 95 KDa. It has a Km value of 0.6 mM for ATP and is maximally active at pH 9.

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DNA-Independent ATPase Activity of Deinococcus radiodurans RecA Protein Is Activated by High Salt (고농도 염에 의한 Deinococcus radiodurans RecA 단백질의 DNA 비의존성 ATPase 역가의 활성화)

  • Kim, Jong-Il
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.313-318
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    • 2010
  • Deinococcus radiodurans RecA protein, when bound to DNA, exhibits a DNA-dependent ATPase. In the absence of DNA, the rate of RecA protein-promoted ATP hydrolysis drops 1,000-fold under the physiological concentrations of salt. This DNA-independent activity can be stimulated to levels approximating those observed with DNA by adding high concentrations (approximately 1.6 M) of a wide variety of salts. This effect was characterized by varying salt concentration and comparing the effects of different ion types. The higher concentrations of salt stimulated the ATP hydrolysis by RecA protein in the absence of DNA. At 1.6 M chloride, the observed stimulation showed the following cation trend $K^+{\geq}Na^+$ > $NH_4^+$ and the following anion sequence was observed: $glutamate^- \; > \; C1^- \;> \; acetate^-\; > \;PO_4^-$ at 1.6 M $K^+$. The catalytic properties of the salt-stimulated ATP hydrolysis reaction was optimal between pH 7.0 and 8.0, which was similar to the double stran nded DNA-dependent ATPase activities of Deinococcus radiodurans RecA protein. In the absence of DNA the active species for ATP hydrolysis by RecA protein was shown to be an aggregate of three RecA protein molecules.

Changes in the Physicochemical Properties of Spent-hen Meat during Cold and Frozen Storage (산란노계육(産卵老鷄肉)의 냉장 및 동결저장 중 물리화학적 특성 변화)

  • Gong, Yang-Sug;Moon, Yoon-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.3
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    • pp.55-61
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    • 1987
  • This study was carried out to investigate the changes in the pH, extractability of protein, ATPase activity of myofibrillar protein, myofibrillar fragmentation, freezing loss and drip loss during storage at $4^{\circ}C$ and $-20^{\circ}C$ in breast and leg muscle of spent-hen meat. pH values ill pectoral and leg muscle were lowest ell tile 1st day and 1st week during cold and frozen storage, respectively. The extractabilities of myofibrillar proteins were increased graduall during cold storage and were highest on the 1st week during frozen storage, The $Mg^{2+}-ATPase$ activities of myofibrillar proteins were highest on the 1st day and 1st week during cold and frozen storage, respectively. The myofibrillar fragmentations were greatly changed on the 1st day during cold storage and 1st week during frozen storage. Freezing losses and drip losses were increased gradually during frozen storage. pH values in breast muscle were lower than those of leg muscle, and the extractabilities, $Mg^{2+}-ATPase$ activities, fragmentations of myofibrillar proteins, and drip losses in breast muscle were higher than those of leg muscle during storage, but the patterns of the changes in both muscles were similar during storage.

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Antifungal Activity of Medium-Chain ($C_{6}-C_{13}$) Alkenals against, and Their Inhibitory Effect on the Plasma Membrane $H^{+}$-ATPase of Saccharomyces cerevisiae

  • Lee, Jae-Ran;Lee, Sang-Hwa;Kubo, Isao;Hong, Soon-Duck
    • Journal of Microbiology and Biotechnology
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    • v.8 no.3
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    • pp.197-202
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    • 1998
  • Aliphatic alkenals having 6 to 13 carbons were evaluated for antifungal activity against Saccharomyces cerevisiae. The activity was gradually increased with chain length, e.g., (E)-2-decenal and (E)-2-undecenal exhibited maximum potency, while (E)-2-dodecenal and (E)-2-tridecenal were completely inactive. Alkenals showed increasing inhibitory activity with chain length, as in the case of antifungal activity, towards glucose-induced medium acidification by the plasma membrane $H^+$-ATPase of S. cerevisiae. The group including (E)-2-nonenal, (E)-2-decenal, and (E)-2-undecenal exhibited maximum potency, but the potency of (E)-2-dodecenal and (E)-2-tridecenal demonstrated a sudden drop with respect to the former group. (E)-2-Nonenal revealed dose-responsive inhibition to the medium acidification and inhibited over 90% at a concentration of 1.25 mM ($175.3{\mu}g$/ml). In contrast to (E)-2-undecenal whose inhibitory efficiency increased with incubation time, inhibition by (E)-2-dodecenal was reversed with time. Of the tested alkenals, (E)-2-heptenal and (E)-2-octenal most highly inhibited ATP hydrolytic activity by the plasma membrane $H^+$ ATPase, while (E)-2-heptenal at 10 mM ($1121.8{\mu}g$/ml) showed an inhibitory efficacy of 93.2%.

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Purification and Sidedness of Sarcolemma from Canine Ventricle (개 심실 형질막의 분리 및 그 방향성에 관한 연구)

  • 이신웅;구정옥;이정수
    • YAKHAK HOEJI
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    • v.30 no.1
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    • pp.31-41
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    • 1986
  • Sarcolemmal membrane fraction from canine ventricle was isolated from the discarded pellet after the first homogenization in the isolation procedure of sarcoplasmic reticulum (Method 1) and the protein yield, purity, and sidedness of this preparation were compared to those of sarcolemmal fraction prepared by method of Lee et al. (Method 2) and a slight modification of original protocol of Jones et al. (Method 3). Method 1 differed from Method 2 essentially only in that vigorous homogenization was carried out by omnimixer and homogenization medium containing 30mM Tris-maleate was used in the first step. The sarcolemmal fraction was enriched from 45 to 50 and 29-fold in [$^3H$] ouabain, [$^3H$] DHA, [$^3H$] QNB binding and $Na^+$, $K^+$-ATPase activity, respectively, compared to homogenate. Total $Na^+$, $K^+$-ATPase activity of highly sarcolemma enriched fraction was 144.6$\pm$16.4$\mu\textrm{mol}$ Pi/mg protein/hr, which was about 85%, of total ATPase activity, and the yield of the preparation was 15.7 mg protein per 100g of starting ventricular tissue. The sarcolemmal preparation supported $^{45}Ca^{2+}$-uptake in the presence of ATP but this uptake was not dependent on oxalate. Sarcolemmal $Na^+$, $K^+$-ATPase activity and detectable [$^3H$] ouabain binding were increased about 32% and 35%, respectively, by pretreatment of sarcolemmal fraction with optimal concentration of sodium dodecylsulfate (0.3-0.4mg/mg protein), suggesting that this preparation contained about 24% of sealed rightside-out vesicles, 26% of sealed inside-out vesicles, and 5001o of freely permeable (leaky) form. This procedure showed the highest protein yield and leaky population, compared to Method 2 and 3. On the other hand, sarcolemmal fraction prepared by Method 2 and 3 showed low value in protein yield but comtained high population of inside-out (46%) and rightside-out (49%) vesicles, respectively, compared to present procedure (Method 1). The results indicate that vigorous homogenization decreases the population of sealed sarcolemmal vesicles but increases the sarcolemmal protein yield per gram tissue and that this procedure is available for further purification of sarcolemmal fraction and for the receptor binding study of sarcolemma.

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Acid Stress Response of Lactobacillus rhamnosus GG in Commercial Yogurt (발효유제품에서 Lactobacillus rhamnosus GG의 생육 특성)

  • Bang, Miseon;Jeong, Anna;Park, Dong-June;Lim, Kwang-Sei;Oh, Sejong
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.1
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    • pp.83-91
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    • 2015
  • Yogurt is a product of the acidic fermentation of milk, which affects the survival of lactic acid bacteria (LAB). The aim of this present study was to examine the survival and acid stress response of Lactobacillus rhamnosus GG to low pH environment. The survival of LAB in commercial yogurt was measured during long-term storage. The enumeration of viable cells of LAB was determined at 15-day intervals over 52-weeks at $5^{\circ}C$. L. acidophilus, L. casei, and Bifidobacterium spp. showed low viability. However, L. rhamnosus GG exhibited excellent survival throughout the refrigerated storage period. At the end of 52-weeks, L. rhamnosus GG survived 7.0 log10 CFU/mL. $F_0F_1$ ATPase activity in L. rhamnosus GG at pH 4.5 was also evaluated. The ATPase activities of the membranes were higher when exposed at pH 4.5 for 24 h. The survival of L. rhamnosus GG was attributable to the induction in $F_0F_1$ ATPase activity. In addition, the mRNA expression levels of acid stress-inducible genes at low pH were investigated by qRT-PCR. clpC and clpE genes were up-regulated after 1 h, and atpA and dnaK genes were up-regulated after 24 h of incubation at pH 4.5. These genes could enhance the survival of L. rhamnosus GG in the acidic condition. Thus, the modulation of the enzymes or genes to assist the viability of LAB in the low pH environment is thought to be important.

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Physiological Changes of Saccharomyces cerevisiae by High Voltage Pulsed Electric Field Treatments (고전압 펄스 전기장 처리에 의한 Saccharomyces cerevisiae의 생리적 변화)

  • Park, Hee Ran;Yoon, So Jung;Park, Han-Sul;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.45 no.5
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    • pp.590-597
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    • 2013
  • High voltage pulsed electric fields (PEF) treatment is a promising non-thermal processing technology that can replace or partially substitute for thermal processes. The aim of this research was to investigate the microbial inactivation mechanisms by PEF treatment in terms of physiological changes to Saccharomyces cerevisiae. PEF was applied at the electric field strength of 50 kV/cm, treatment time of 56 ${\mu}s$ and temperature of $40^{\circ}C$. The microbial cells treated with PEF showed loss of salt tolerance on the cell membrane and collapse of the relative pH gradient on in-out of cells. Cell death or injury resulted from the breakdown of homeostasis, decreased $H^+$-ATPase activity, and loss of glycolysis activity.

Different Distribution of the ${alpha}_{2},Na^+,K^+-ATPase lsoform between Rat Atria and Ventricles$

  • Lee, Jeung-Soo;Lee, Shin-Woong;Wallick, Earl-T
    • Archives of Pharmacal Research
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    • v.19 no.5
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    • pp.381-385
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    • 1996
  • Rat ventricles respond with a biphasic positive inotropic effect to ouabain, low-dose and high-dose effects but rat atria with only a monophasic high dose effect. In an effect to understand the difference in response to ouabain of two tissues between rat atria and ventricles the levels of the $a_{2}$ -isoform of the $Na^{+}$, $K^{+}$-ATPase which has higher affinity for ouabain than the $a_{1}$-iso-form were determined by a $[^{3}H]$ouabain binding assay. The yield of protein per gram wet weight was about 47 mg for atria and 100 mg for ventricles. The $K_{d}$ values of ouabain for the high-affinity ouabain binding site $(a_{2} -isoform)$ were nearly the same (230 nM) in the atria and ventricles. However, the numbers of the $a_{2}$-isoform $(B_{max})$ per mg protein were approximately half in the atria. When the binding data were expressed in unit per gram tissue wet weight, the numbers of $a_{2}$ -isoform in the atria was about 25% of that in the ventricles. THese results demonstrate that the $a_{2}$ -isoform of the $Na^{+}$, $K^{+}$-ATPase in the rat atria could be detected by $[^{3}H]$ouabain binding assay and the levels of this isoform are too low to show the low-dose effect of ouabain.

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Structural Analysis of Lamellar Plane in Three-Dimensional Crystal of $Ca^{2+}-ATPase$ ($Ca^{2+}-ATPase$ 3차원 결정의 Lamellar면 구조분석)

  • Cheong, Gang-Won
    • Applied Microscopy
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    • v.27 no.2
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    • pp.111-120
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    • 1997
  • Cylinder-like crystals of $Ca^{2+}-ATPase$ provide views parallel to the lamellar plane, from which parameters of lamellar stacking can be directly measured. These parameters were measured using different preparation methods. Assuming that molecular packing is the same, data from lamellar plane could supplement those obtained by tilting large, thin plate-like crystals. However, base on data obtained .by electron microscopy and x-ray powder patterns, the plate-like crystal may have another scheme for stacking the lamellar. The projection map (h, 0, 1) from cylinder-like crystals using cryoelectron microscopy suggest the lamellar spacing can be variable.

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