• The Korean Journal of Physiology
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• v.29 no.1
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• pp.69-80
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• 1995

Renal proximal tubular hypertrophy and hyperfunction are known to be early manifestations of experimental and human diabetes. As the hypertrophy and hyperfunction have been suggested to be central components in the progression to renal failure, an understanding of their underlying causes is potentially important for the development of therapy. A primary rabbit kidney proximal tubule cell culture system was utilized to evaluate the possibility that the renal proximal tubular hypertrophy and hyperfunction observed in vivo in diabetes mellitus, can be attributed to effects of elevated glucose levels on membrane transport systems. Primary cultures of rabbit proximal tubules, which achieved confluence at 10 days, exhibited brush-border characteristics typical of proximal tubular cells. Northern analysis indicated $2.2{\sim}2.3$ and 2.0 kb Na/glucose cotransporter RNA species appeared in fresh and cultured proximal tubule cells after confluence, repectively. The cultured cells showed reduced Na/glucose cotransporter activity compared to fresh proximal tubules. Primary cultured proximal tubule cells incubated in medium containing 20 mM glucose have reduced ${\alpha}-MG$ transport compared to cells grown in 5 mM glucose. In the proximal tubule cultures incubated in medium containing 5 mM or 20 mM glucose, phlorizin at 0.5 mM inhibited 0.5 mM ${\alpha}-MG$ uptake by 84.35% or 91.85%, respectively. The uptake of 0.5 mM ${\alpha}-MG$ was similarly inhibited by 0.1 mM ouabain (41.97% or 48.03% inhibition was observed, respectively). In addition, ${\alpha}-MG$ uptake was inhibited to a greater extent when $Na^{+}$ was omitted from the uptake buffer (81.86% or 86.73% inhibition was observed, respectively). In cell homogenates derived from the primary cells grown in 5 mM glucose medium, the specific activity of the Na/K-ATPase $(6.17{\pm}1.27\;{\mu}mole\;Pi/mg\;protein/hr)$ was 1.56 fold lower than the values in cell homogenates treated with 360 mg/dl D-glucose, 20 mM $(9.67{\pm}1.22\;{\mu}mole\;Pi/mg\;protein/hr)$. Total $Rb^{+}$ uptake occurred at a significantly higher rate (1.60 fold increase) in primary cultured rabbit kidney proximal tubule cell monolayers incubated in 20 mM glucose medium $(10.48{\pm}2.45\;nM/mg\;protein/min)$ as compared with parallel cultures in 5 mM glucose medium. $Rb^{+}$ uptake rate in 5 mM glucose medium was reduced by 28% when the cultures were incubated with 1 mM ouabain. The increase of the $Rb^{+}$ uptake by rabbit kidney proximal tubule cells in 20 mM glucose could be attributed primarily to an increase in the rate of ouabain-sensitive $Rb^{+}$ uptake $(5\;mM\;to\;20\;mM;\;4.68{\pm}0.85\;to\;8.38{\pm}1.37\;nM/mg\;protein/min)$. In conclusion, the activity of the renal proximal tubular Na,K-ATPase is elevated in high glucose concentration. In contrast, the activity of the Nafglucose cotransport system is inhibited.

• The Korean Journal of Physiology and Pharmacology
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• v.17 no.4
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• pp.275-281
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• 2013

Astrocytes are reported to have critical functions in ischemic brain injury including protective effects against ischemia-induced neuronal dysfunction. Na-K ATPase maintains ionic gradients in astrocytes and is suggested as an indicator of ischemic injury in glial cells. Here, we examined the role of the Na-K ATPase in the pathologic process of ischemic injury of primary cultured astrocytes. Chemical ischemia was induced by sodium azide and glucose deprivation. Lactate dehydrogenase assays showed that the cytotoxic effect of chemical ischemia on astrocytes began to appear at 2 h of ischemia. The expression of Na-K ATPase ${\alpha}1$ subunit protein was increased at 2 h of chemical ischemia and was decreased at 6 h of ischemia, whereas the expression of ${\alpha}1$ subunit mRNA was not changed by chemical ischemia. Na-K ATPase activity was time-dependently decreased at 1, 3, and 6 h of chemical ischemia, whereas the enzyme activity was temporarily recovered to the control value at 2 h of chemical ischemia. Cytotoxicity at 2 h of chemical ischemia was significantly blocked by reoxygenation for 24 h following ischemia. Reoxygenation following chemical ischemia for 1 h significantly increased the activity of the Na-K ATPase, while reoxygenation following ischemia for 2 h slightly decreased the enzyme activity. These results suggest that the critical time for ischemia-induced cytotoxicity of astrocytes might be 2 h after the initiation of ischemic insult and that the increase in the expression and activity of the Na-K ATPase might play a protective role during ischemic injury of astrocytes.

• Fisheries and Aquatic Sciences
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• v.2 no.2
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• pp.161-166
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• 1999

The rigor-mortis progress of cultured olive flounder spiked at the brain started much faster than that of wild one. They attained full rigor state after 30 hrs at $0^{\circ}C$, 36 hrs at $5^{\circ}C$ and 50 hrs at $10^{\circ}C$ in the cultured flounder, while after 36 hrs at $0^{\circ}C$, 50 hrs at $5^{\circ}C$, and 60 hrs at $10^{\circ}C$ in the wild. ATP concentration in the muscle was around $5.9\mu mol/g$ for wild and $6.2\mu mol/g$ for cultured flounder. ATP breakdown progressed rapidly in $0^{\circ}C$ samples, followed by $5^{\circ}C$ and $10^{\circ}C$ samples. $Mg^{2+}$-ATPase activity of myofibrillar protein in the presence of 0.25mM CaCb was higher in cultured myofibri1lar protein than in wild one. $Mg^{2+}$-ATPase activities of myofibrillar protein increased during storage in samples stored at $0^{\circ}C$ and $5^{\circ}C$ while decreased in samples stored at $10^{\circ}C$. The level of breaking strength of muscle immediately after death was higher in the wild muscle than in the cultured muscle. The breaking strength reached maximum level at 10 hrs after death in both samples.

• Journal of Radiation Protection and Research
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• v.20 no.2
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• pp.71-78
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• 1995

To develop methods to reduce radiation risk and apply such knowledge to improvement of radiation protection, the effects of cobaltous chloride known as bioantimutagen on the function of E. coli RecA protein involved in the repair of DNA damage were examined. The results demonstrated two distinct effects of cobaltous chloride on the RecA protein function necessary for the strand exchange reaction. Cobaltous chloride enhanced the ability of RecA protein to displace SSB protein from single-stranded DNA and the duplex DNA-dependent ATPase activity. RecA protein was preferentially bound with UV-irradiated supercoiled DNA as compared with nonirradiated DNA The binding of RecA protein to UV-irradiated supercoiled DNA was enhanced in a dose-dependent manner. It is likely that studies on the factors affecting repair efficiency and the DNA repair proteins may provide information on the repair of ionizing radiation-induced DNA damage and the mechanism for DNA radioprotection.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.574-580
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• 2019

ATP drives the conformational change of the group II chaperonin from the open lid substrate-binding conformation to the closed lid conformation to encapsulate an unfolded protein in the central cavity. It is thought that the folding activity of group II chaperonin is strongly correlated with the ATP-dependent conformational change ability. In order to confirm the dependence of the reaction temperature and ATP concentration of PhCpn, the ATPase activities were measured under different reaction temperatures and ATP concentrations. The maximal ATPase activity of PhCpn was observed at 80℃ and 3 mM ATP concentration. As a result of ATPase activity according to the type of salt ions, the highest activity was observed at 300 mM LiCl among the univalent cations and 5 mM MgCl₂ among the divalent cations, respectively. The values of Km and Vmax for ATP substrate were estimated as 2.17 mM and 833.3 μM/min, respectively. This results provide the enzymatic information of PhCpn when the prolonged and high activities of pharmaceutical and industrial proteins (or enzymes), by using chaperonin molecules, are required.

• The Korean Journal of Zoology
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• v.28 no.1
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• pp.31-43
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• 1985

The $(Ca^{2+}+Mg^{2+})$-ATPase has been purified homogeneously from sarcoplasmic reticulum of rat skeletal muscle by sucrose density gradient centrifugation. The purified enzyme has a molecular weight of 115,000 as judged by polyacrylamide gel electrophoresis in the presence of sodium dedecyl sulfate, and therefore has the same size of the enzyme in rabbit and chick skeletal muscle. $Ca^{2+}, Mg^{2+}, Fe^{2+}, Co^{2+}, and Mn^{2+}$ at 50 $\\muM$ show stimulatory effect on the ATP-ase, while $Zn^{2+}, Cu^{2+}, and Hg^{2+}$ inhibit it at the same concentration. The ATPase activity is insensitive to antimalarial drugs such as quinine and quinacrine, but is sensitive to inhibition by p-hydroxymecurie benzoate and phenylmethylsulfonylfluoride. The enzyme has optimum pH of 6 to 7 and Km value for ATP is estimated to be 98 $\\muM$. Thus, a number of biochemical properties of this enzyme appear to be different from those of the enzyme that have been isolated from rabbit skeletal muscle. The $(Ca^{2+}+Mg^{2+})$-ATPase appears to be selectively degraded in microsomal fraction. The activity of metalloendoprotease is evident in the microsomal preparation when assayed by radioactively labeled protein substrate, such as $^{3}H-casein and $^{125}I$-insulin. However, it is presently unclear whether the metalloendoprotease is responsible for the degradation of the $(Ca^{2+}+Mg^{2+})$-ATPase.

• 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.

• The Korean Journal of Physiology and Pharmacology
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• v.12 no.6
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• pp.331-336
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• 2008

The present study was aimed to determine whether there is an altered regulation of tubular transporters in gentamicin-induced nephropathy. Sprague-Dawley male rats ($200{\sim}250\;g$) were subcutaneously injected with gentamicin (100 mg/kg per day) for 7 days, and the expression of tubular transporters was determined by immunoblotting and immunohistochemistry. The mRNA and protein expression of OAT was also determined. Gentamicin-treated rats exhibited significantly decreased creatinine clearance along with increased plasma creatinine levels. Accordingly, the fractional excretion of sodium increased. Urine volume was increased, while urine osmolality and free water reabsorption were decreased. Immunoblotting and immunohistochemistry revealed decreased expression of $Na^+/K^+$-ATPase, NHE3, NBC1, and AQP1 in the kidney of gentamicin-treated rats. The expression of OAT1 and OAT3 was also decreased. Gentamicin-induced nephropathy may at least in part be causally related with a decreased expression of $Na^+/K^+$-ATPase, NHE3, NBC1, AQP1 and OAT.

• Food Science of Animal Resources
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• v.27 no.4
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• pp.457-462
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• 2007

Effects of sonication on water-solubilization of myofibril from breast muscle of spent hen effects investigated in this study. To evaluate effect of salt concentration and pH, salt concentration was varied with range from 0.1 to 0.8 M, and pH was varied with range from 6.0 to 8.0. Solubility, SDS-PAGE, viscosity and ATPase activity of sonicated myofibril were measured. Solubility of myofibrillar protein containing 0.1 M NaCl at pH 8.0 after sonication was above 90%. Main components of soluble protein by SDS-PAGE were myosin heavy chain and actin. That is, it indicated breaking of myofibril structure by sonication. Also, viscosity of soluble protein increased, but Ca- and Mg-ATPase activities decreased by increasing sonication time. From these results, we concluded that most of myofibrillar proteins were denatured by sonication.

• Proceedings of the Zoological Society Korea Conference
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• 1996.10a
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• pp.180.1-180
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• 1996

No Abstract, See Full Text