• Korean Journal of Environmental Agriculture
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• v.21 no.2
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• pp.102-108
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• 2002

In order to investigate the mechanism of growth inhibition by salt stress, lettuces were grown hydroponically in three different nutrient solutions, normal and 30 mM or 50 mM $KNO_3$-added nutrient solutions, and the electrical conductivities of these solutions were 1.0, 4.5, and 6.5 dS/m, respectively. The activities of plasma and vacuolar $H^+$-ATPases in the root tissue of lettuce were measured by specific inhibitors, 100 ${\mu}M$ vanadate and 50 mM $NO_3^-$, respectively. Microsomal ATPase activity of lettuce grown in the normal nutrient solution was $356\pm1.5$ nmol/min/mg protein. When lettuces were grown in 30 mM and 50 mM $KNO_3$-added nutrient solutions, total activities of microsomal ATPases were increased by 1.6 and 1.9 times, respectively, and the increases were mainly mediated by vacuolar $H^+$-ATPase. These results show that lettuces adapt themselves to salt-stressed condition by increasing the activities of $H^+$-ATPases. Effects of various heavy metal ions were investigated on the microsomal ATPases and various metal ions at 100 $\mu M$ inhibited the activities by 10$\sim$25%. $Cu^{2+}$ showed the highest inhibitory effect on the vacuolar $H^+$-ATPase. These results suggest that lettuce increases the activities of root ATPases, specially that of vacuolar $H^+$-ATPase, in salt-stressed growth conditions and $Cu^{2+}$ could be a useful tool to control the activity of vacuolar $H^+$-ATPase.

• Journal of Bio-Environment Control
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• v.18 no.2
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• pp.160-165
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• 2009

This study was undertaken the effect of high temperature and high humidity on protein expression and especially plasma membrane (PM) $H^{+}ATPase$ of umbel with flowers of early cultivar 'Shinsunhwang' and intermediate cultivar 'Maebsihwang' of onion (Allium cepa L.). There were no visible any difference on the protein pattern from before flowering stage to full flowering stage of two onion cultivars, however, seed set stages were revealed induced/deduced protein patterns. At day 18, protein expression pattern of the high temperature and high humidity treatments of two cultivars was significantly reduced compared to controls. Furthermore, various protein expression of the high temperature treatment was more reduced compared to high humidity treatment. PM $H^{+}ATPase$ expression of the control plants of two onion cultivars was clearly shown, but was not detectable under high temperature treatment of the two onion cultivars using western blot analysis. PM $H^{+}ATPase$ expression of the high humidity treatment was faintly detected intermediate cultivar 'Maebsihwang', not early cultivar 'Shinsunhwang'. These results indicate that protein expression pattern and PM $H^{+}ATPase$ under high temperature treatment was considered to be more damaged compared to high humidity.

• Animal cells and systems
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• v.8 no.4
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• pp.307-312
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• 2004

Fast kinetics of transient pH changes and difference spectrum formation have been investigated following mixing of ADP/ATP with partially purified plasma membrane PM-ATPase of the pathogenic yeast Candida albicans in the presence of five nutrients: glucose, glutamic acid, proline, lysine, and arginine and two analogs of glucose: 2-deoxy D-glucose and xylose. Average $H^+$- absorption to release ratio, indicative of population of ATPase undergoing complete hydrolytic cycle, was found to be 0.27 for control. This ratio varied between 0.25 (proline) to 0.36 (arginine) for all other compounds tested, except for glucose. In the presence of glucose, $H^+$- absorption to release ratio was exceptionally high (0.92). While no UV difference spectrum was observed with ADP, mixing of ATP with ATPase led to a large conformational change. Exposure to different nutrients restricted the magnitude of the conformational change; the analogs of glucose were found to be ineffective. This suppression was maximal in the case of glucose (80%); with other nutrients, the magnitude of suppression ranged from 40-50%. Rate of $H^+$- absorption, which is indicative of E~P complex dissociation, showed positive correlation with suppression of conformational change only in the case of glucose and no other nutrient/analog. Mode of interaction of glucose with plasma membrane $H^+$-ATPase thus appears to be strikingly distinct compared to that of other nutrients/analogs tested. The results obtained lead us to propose a model for explaining glucose stimulation of plasma membrane $H^+$-ATPase activity.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.1
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• pp.72-79
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• 2011

Here we report a simple screening system using hematoxylin staining (HS) of the root apex. It allowed rapid classification into different aluminum (Al) tolerance from 65 cultivars within one week. Using this system, we selected the most Al-tolerant (Jayae-2) and-sensitive (Pum-2) barley (Hordeum vulgare L.) The results show that the different responses in Al-induced growth inhibition, Al accumulation and expression of plasma membrane (PM) $H^+$-ATPase in root apices of selected two cultivars. It showed strongly Al-induced growth inhibition in a dosedependant manner only in Pum-2 but not in Jayae-2. Aluminum accumulation in root apices (10 mm) was significantly higher in Pum-2 only. The $H^+$-ATPase expression of PM vesicles by western blotting was decreased in Pum-2 but not in Jayae-2 treated with $20{\mu}M$ Al for 24 h. These finding indicate to screen from our system is rapid and reliable and to sustain the expression of PM $H^+$-ATPase at translational level is an important role in root growth as affected by Al.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.2
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• pp.73-78
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• 1992

To investigate the desirable index for evaluation of frozen surimi's grades, the relationship between indices for grading of surimi, such as ATPase activity $(Ca^{2+}-,\;Mg^{2+}-\;and\;EDTA-)4, solubility, viscosity and K-value of frozen surimi and jelly strength of kamaboko was studied. The myofibrillar $Ca^{2+}-ATPase$ activity and solubility from frozen surimi of grades SA, FA, A, RA and B gave values of $1.184\pm0.2,\;0.956\pm0.14,\;0.766\pm/0.07,\;0.453\pm0.07\;and\;0.227\pm0.08$(umoles Pi/min/mg) for $Ca^{2+}-$ATPase activity and $93.19\pm5,\;84.62\pm4,\;70.63\pm5,\;41.21\pm4\;and\;32.82\pm4(\%)$ for solubility, respectively. Therefore, the myofibrillar $Ca^{2+}-$ATPase activity and solubility of surimi were closely related to the jelly strength of kamaboko from same material, as the correlation coefficient were 0.9584 and 0.9849, respectively. K-value, the index of freshness, was related to the jelly strength of frozen surimi as the correlation coefficient 0.9053 and shown as SA $15.67\pm1.4,\;FA\;14.94\pm 3,\;A\;28.00\pm5,\;RA\;32.16\pm3\;and \;B\;48.68\pm 5(\%)$. $Mg^{2+}-$ and EDTA-ATPase activity and viscosity were not related to the jelly strength. The $Ca^{2+}-$ATPase activity and solubility were found to be useful index for evaluating the quality of frozen surimi.

• Ecology and Resilient Infrastructure
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• v.10 no.3
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• pp.64-72
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• 2023

Biopolymer is a versatile material used in food processing, medicine, construction, and soil reinforcement. 𝛽-glucan is one of the biopolymers that improves the soil water content and ion adsorption in a drought or toxic metal contaminated land for plant survival. We analyzed drought stress damage reduction in sweet potatoes (Ipomoea batatas L. cv. Sodammi) by measuring the growth and major protein expression and activity under 𝛽-glucan soil amendment. The result showed that sweet potato leaf length and width were not affected by drought stress for 14 days, but sweet potatoes grown in 𝛽-glucan-amended soil showed an effect in preventing wilting caused by drought in phenotypic changes. Under drought stress, sweet potato leaves did not show any changes in electrolyte leakage, but the relative water content was higher in sweet potatoes grown in 𝛽-glucan-amended soil than in normal soil. 𝛽-glucan soil amendment increased the expression of plasma membrane (PM) H⁺-ATPase, but it decreased the aquaporin PIP2 (plasma membrane intrinsic protein 2) in sweet potatoes under drought stress. Moreover, water maintenance affected the PM H⁺-ATPase activity, which contributed to tolerance under drought stress. These results indicate that 𝛽-glucan soil amendment improves the soil water content during drought and affects the water supply in sweet potatoes. Consequently, 𝛽-glucan is a potential material for maintaining soil water contents, and analysis of the major PM proteins is one of the indicators for evaluating the biopolymer effect on plant survival under drought stress.

• Plant Biotechnology Reports
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• v.3 no.4
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• pp.293-299
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• 2009

Auxin-binding protein 57 ($ABP_{57}$), a soluble auxin-binding protein, acts as a receptor to activate plasma membrane (PM) $H^+-ATPase$. Here, we report the cloning of abp57 and the biochemical characterization of its protein expressed in E. coli. The analysis of internal amino acid sequences of $ABP_{57}$ purified from rice shoots enabled us to search for the corresponding gene in protein DB of NCBI. Further BLAST analysis showed that rice has four abp57-like genes and maize has at least one homolog. Interestingly, Arabidopsis seems to have no homolog. Recombinant $ABP_{57}$ expressed in E. coli caused the activation of PM $H^+-ATPase$ regardless of the existence of IAA. Scatchard analysis showed that the recombinant protein has relatively low affinity to IAA as compared to natural $ABP_{57}$. These results collectively support the notion that the cloned gene is responsible for $ABP_{57}$.

• 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.

• Biomolecules & Therapeutics
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• v.23 no.1
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• pp.53-59
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• 2015

In this study, we investigated the inhibitory activities on gastritis and gastric ulcer using liriodendrin which is a constituent isolated from Kalopanax pictus. To elucidate its abilities to prevent gastric injury, we measured the quantity of prostaglandin $E_2$ ($PGE_2$) as the protective factor, and we assessed inhibition of activities related to excessive gastric acid be notorious for aggressive factor and inhibition of Helicobacter pylori (H. pylori) colonization known as a cause of chronic gastritis, gastric ulcer, and gastric cancer. Liriodendrin exhibited higher $PGE_2$ level than rebamipide used as a positive control group at the dose of $500{\mu}M$. It was also exhibited acid-neutralizing capacity (10.3%) and $H^+/K^+$-ATPase inhibition of 42.6% ($500{\mu}M$). In pylorus-ligated rats, liriodendrin showed lower volume of gastric juice ($4.38{\pm}2.14ml$), slightly higher pH ($1.53{\pm}0.41$), and smaller total acid output ($0.47{\pm}0.3mEq/4hrs$) than the control group. Furthermore liriodendrin inhibited colonization of H. pylori effectively. In vivo test, liriodendrin significantly inhibited both of HCl/EtOH-induced gastritis (46.9 %) and indomethacin-induced gastric ulcer (46.1%). From these results, we suggest that liriodendrin could be utilized for the treatment and/or protection of gastritis and gastric ulcer.

• Korean Journal of Veterinary Research
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• v.49 no.1
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• pp.17-22
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• 2009

The effect of Bu-Zhong-Yi-Qi-Tang (BZYQT) on the changes of ultraviolet (UV) light B radiation-induced apoptotic sunburn cell (SBC) and epidermal ATPase-positive dendritic cell (DC) in SKH1- hr or ICR mouse were investigated. The mice were treated with UVB (200 mJ/$cm^2$) and were sacrificed 24 h later. BZYQT (50 mg/kg of body weight) or vehicle (saline) was given i.p. at 36 and 12 h before irradiation, and 30 min after irradiation or BZYQT cream (0.2%) or cream base (vehicle) was topically treated at 24 h and 15 min before irradiation, and immediately after irradiation. The skin of SKH1-hr mouse prepared from the back of untreated mice exhibited about 0.3 SBC/cm length of epidermis, and 24 h after UV irradiation, the applied areas show an increased number of SBCs. But the frequency of UVB-induced SBC formation was reduced by intraperitoneal injection of BZYQT extract (p < 0.01). The numbers of DC in normal ICR mouse were 628.00 ${\pm}$ 51.56 or 663.20 ${\pm}$ 62.58 per $mm^2$ of ear epidermis. By 1 day after UVB treatment, the number of ATPase-positive cells/$mm^2$ were decreased by 39.0% or 27.1% in i.p. or topical application group with vehicle. Treatment of BZYQT was associated with increase of 33.9% in i.p. group (p < 0.05) or 2.7% in topical application group in the number of ATPase positive cells compared with the irradiation control group. The results presented herein that BZYQT administration could reduce the extent of skin damages produced by UVB.