• 한국미생물·생명공학회지
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• 제33권3호
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• pp.184-188
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• 2005

HRF는 Na,K-ATPase의 ${\alpha}$ subunit에 결합하여 이의 활성을 저해하는 것으로 알려져 있으며, PKC에 의해 Ser98 잔기가 인산화 될 수 있다는 것을 anti-HRFpS98 항체와 HRF S98A mutant를 이용한 실험으로 확인할 수 있었다. 또한 $^{86}Rb^{+}-uptake$ assay 실험에서 HRF의 serine 98 잔기의 탈인산화는 Na,K-ATPase의 활성에 약간의 영향을 미치는 것으로 미루어 PKC에 의해 인산화되는 98 serine 잔기가 Na,K-ATPase 활성 저해에 큰 영향을 미치지 않는 것으로 보인다.

• The Korean Journal of Physiology
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• 제1권2호
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• pp.141-150
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• 1967

The present investigation was initially undertaken to see if there exists $Na^+-K^+$ activated ATPase in the microsome fraction of the kidney. Having confirmed the presence of such an enzyme, further attempts have been made to characterize its nature and the following conclusions were obtained: (1) The ATPase activity was greatest at the $Na^+$ concentration of 100 mM as well as at $K^+$ concentration of 10 mM. Moreover, the ATPase activity was found to be depressed by $Ca^{++}$ in the presence of $Mg^{++}$. (2) While the ATPase activity was depressed by Ouabain, the magnitude of inhibition was greater in the Na medium than in the K medium. (3) NaCN augmented the ATPase activity whereas NaF and IAA depressed it. On the other hand, DNP had little influence on the ATPase activity. (4) Diamox, vasopressin and aldosterone had no effect while $HgCl_2$ markedly depressed the ATPase activity These findings indicate that the nature of ATPase isolated from the microsome fraction of the rabbit kidney is quite similar to that from other organs such as the heart and the muscle, although there are certain features specific to the type of organs.

• 환경생물
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• 제34권2호
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• pp.97-106
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• 2016

$Na^+/K^+$-ATPase is a membrane protein and plays a key role in osmotic regulation in living organisms. In the present study, a cDNA sequence encoding the $Na^+/K^+$-ATPase alpha subunit from the monogonont rotifer, Brachionus koreanus was cloned by rapid amplification of cDNA ends technique. To investigate the role of this enzyme in osmotic stress, enzymatic activities of $Na^+/K^+$-ATPase were measured after exposure to different salinities for 48 h. The full-length Bk $Na^+/K^+$-ATPase cDNA was 3069 bp-long, encoding a 1022-amino acid polypeptide. Bk $Na^+/K^+$-ATPase possesses eight membrane spanning regions and five conserved domains. Phylogenetic analysis showed that Bk $Na^+/K^+$-ATPase had high identity with those of other species, and was closely clustered with other Brachionus sp. These findings indicate that this protein was conserved both structurally and functionally. B. koreanus $Na^+/K^+$-ATPase activity was stimulated in both hyposaline (6 psu) and hypersaline (32 psu) conditions, suggesting that this protein may play a role in osmoregulation. This study would provide better understanding of the physiology of B. koreanus and this enzyme may be useful as a molecular marker for evaluation of osmotic stress in aquatic environment.

• 약학회지
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• 제32권1호
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• pp.62-69
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• 1988

We have studied the effect of fasting on $Na^+$, $K^{+}-ATPase$ activities in the rat intestinal mucosa. Rats were fasted for $18{\sim}48hr$. Intestinal microsomal fraction was prepared by the method of Robinson and ATPase activities were determined by the modified method of Fiske and Subbarow. $Na^+$, $K^{+}-ATPase$ activity was not changed after fasting for 18 and 24 hr but significantly decreased after fasting for 48 hr. Fasting over 18 to 48 hr period had no effect on the $Mg^{++}-ATPase$. Thus, it may be concluded that 48 hr fasting has inhibitory effect on rat intestinal absorptive capabilities. In order to study the effect of Ginseng on the $Na^+$, $K^{+}-ATPase$ activities of the small intestine in chronic $K^{+}-depleted$ rats, rats were fed $K^{+}-depleted$ diets for 3 weeks and Ginseng ethanol extracts were administered orally for 3 weeks concomitantly. ATPase activity was measured by the same method as fasting group. $Na^+$, $K^{+}-ATPase$ activity in the $K^{+}-depleted$ diet group was increased and Ginseng ethanol extracts inhibited the increase of enzyme activity induced by $K^{+}-depleted$ diet. Thus, it may be suggested that increase in the intestinal $Na^+$, $K^{+}-ATPase$ activity of chronic $K^{+}-depleted$ group may be due to the compensatory mechanism and administration of Ginseng with $K^{+}-depleted$ diet may be associated with inhibition of increase in the enzyme activity of the $K^{+}-depleted$ group due to the prevention of the $K^+$ loss in the $K^{+}-depletion$.

• The Korean Journal of Physiology
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• 제8권1호
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• pp.67-76
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• 1974

The effect of saponin on the sodium plus potassium activated ATPase activity was studied in the rabbit red cell ghosts and the experiments were also designed to determine the mechanism of action of saponin on the APTase activity. The following results were observed. 1. The ATPase activity of rabbit red cell ghosts is inhibited by low concentration of saponin but increased by high concentration. The activating effect of saponin on the $Na^{+}-K^{+}-ATPase$ activity is inhibited by ouabain but the stimulation of the $Mg^{++}-ATPase$ by high concentration of saponin is not inhibited by ouabain. 2. The activity ratio of $Na^{+}-K^{+}-ATPase$ by high concentration of saponin is decreased by raising the potassium concentration, and is increased by raising the sodium concentration. 3. The ATPase activity is increased by small amounts of calcium but inhibited by larger amounts. The activity ratio of the enzyme by saponin is decreased by raising the calcium concertration 4. The action on the ATPase activity was not related to the amino group of lysine, the hydroxyl group of threonine, the imidazole group of histidine, or the carboxyl group of aspartic acid. 5. The action of saponin on the ATPase activity is due to sulfhydryl group of the enzyme of $Na^{+}-K^{+}-ATPase$.

• 한국물환경학회지
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• 제18권4호
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• pp.395-400
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• 2002

Lead is one of the most toxic metal and is detectable in practically all phases of environment and in all biological system. Transport, industrial and domestic waste products are the main sources of this pollutant. Ingested lead is rapidly absorbed and widely distributed throughout the body, causing extensive tissue damage. In this study, we chose the freshwater decapods Maceobrachium nipponnese as a sensitive indicator organism for environmental pollution. In order to investigate the possibility in use of $Na^+/K^+ATPase$ activity as a biomarker of lead pollution, we tested the acute toxicity of lead to Maceobrachium nipponnese. The $LC_{50}(96hr)$ value for lead in Maceobrachium nipponnese was found to be $446{\mu}g/L$ with the 95% confidence limits. The lead exposure group at $LC_{50}$ showed a significant $Na^+/K^+ATPase$ inhibition, depending on the exposure time. Comparision of several concentrations of lead revealed that the $Na^+/K^+ATPase$ activity in Maceobrachium nipponnese was significantly decreased in a concentration dependent manner. These results suggest that $Na^+/K^+ATPase$ activity in Maceobrachium nipponnese may possibly be used as a biomarker of lead pollution in aquatic ecosystem.

• The Korean Journal of Physiology
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• 제12권1_2호
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• pp.25-34
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• 1978

The action of theobromine on the sodium plus potassium activated ATPase activity In the rabbit red cell membrane has teen investigated and the experiments were also designed to determine the mechanism of action of theobromine on the ATPase activity. The following results were observed. 1. The activity of the NaK ATPase from red fell membrane is stimulated by theobromine, and the concentration of theobromine for maximal activity is about 3mM. 2. The activating effect of theobromine on the ATPase, with a given concentration of potassium in the medium, is increased by raising the sodium concentration but activity ratio is decreased. 3. The activating effect of theobromine on the ATPase, with a given concentration of sodium in the medium. is increased by the raising the potassium concentration but activity ratio is decreased. 4. The NaK ATPase activity is increased by small amounts of calcium but decreased by larger amounts. The activity of the enzyme by theobromine is increased by small amounts of calcium but decreased by larger amounts. 5. The activating effect of theobromine on the ATPase was not related to the hydroxyl group of threonine and imidazole group of histicline. 6. The activating effect of theobromine on the ATPase is due to sulfhydryl group, amino group and carboxyl group of the enzyme of NaK ATPase.

• The Korean Journal of Physiology
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• 제11권1호
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• pp.11-20
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• 1977

The action of pilocarpine on the sodium plus potassium activated ATPase activity in the rabbit red cell membrane has been investigated and the experiments were also designed to determine the mechanism of action of pilocarpine on the ATPase activity. The following results were observed. 1. The activity of the NaK ATPase from red cell membrane is stimulated by pilocarpine, and the concentration of pilocarpine for maximal activity is about 3 mM. The pH optimum for the pilocarpine sensitive component is 8.0. 2. The activating effect of pilocarpine on the ATPase, with a given concentration of sodium .in the medium, is increased by raising the potassium concentration but activity ratio is decreased 3. The activating effect of pilocarpine on the ATPase, with a given concentration of Potassium in the medium, is increased by raising the sodium concentration but activity ratio is decreased 4. The NaK ATPase activity is increased by small amounts of calcium but decreased by 'larger amounts. The activity ratio of the enzyme by pilocarpine is decreased by small amounts .of calcium but decreased by larger amounts. 5. The activating effect of pilocarpine on the ATPase was not related to the sulfhydryl group of cysteine, the hydroxyl group of threonine or the imidazole group of histidine. 6. The activating effect of pilocarpine on the ATPase is due to amino group and carboxyl group of the enzyme of NaK ATPase

• 대한핵의학회지
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• 제32권2호
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• pp.121-128
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• 1998

목적 : 생물학적 성질이 potassium과 유사하고 핵의학분야에서 널리 이용되고 있으며 쉽게 구할 수 있는 T1-201을 이용하여 $Na^+-K^+$ ATPase의 활성도를 측정할 수 있는지를 알아보고자, 배양한 백서 대동맥평활근세포와 사람의 제대동맥 평황근세포에서 $Na^+-K^+$ ATPase의 활성도를 측정하곤 기존의 Rb-86으로 측정한 방법과 비교하였다. 또한 T1-201로 측정한 활성도에 대한 포도당, 인슐린 및 PMA의 영향을 알아보고자 하였다. 대상 및 방법: Sprague-Dawley 백서의 흉부대동맥과 인체태반의 제대동맥에서 얻은 평활근세포를 배양하여 사용하였고. ouabain첨가로 억제되는 Rb-86과 T1-201의 섭취율을 $Na^+-K^+$ ATPase에 의한 섭취율로 계산하여 활성도로 간주하였다. 결과: 배양된 백서대동맥 평활근세포에서 $Na^+-K^+$ ATPase 활성도는 고포도당배양액(22 mM) 상태에서 생리적 농도의 저포도당배양액(5 mM) 상태에 비하여 평균 28%의 저하를 보였으며, 인슐린 100 nM을 첨가하였을 때는 50%의 증가를 보였다. PKC효소를 활성화시키는 PMA는 20%의 증가를 나타내었다. 인체제대동맥의 평활근세포에서도 유사한 변화를 보였다. T1-201로 측정한 $Na^+-K^+$ ATPase의 활성도치의 변화는 Rb-86을 이용한 측정에서도 동일하게 나타났다. 결론: T1-201을 이용하여 측정한 $Na^+-K^+$ ATPase 황성도는 Rb-86을 이용한 측정치와 유사하여, T1-201은 세포막 $Na^+-K^+$ ATPase 활성도의 측정에 사용할 수 있으리라 판단된다. 인슐린과 PKC효소 자극제는 $Na^+-K^+$ ATPase 활성도를 증가시키고, 고농도의 포도당은 활성도를 감소시키는 것으로 사료된다.

• 약학회지
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• 제29권2호
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• pp.76-89
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• 1985

The effects of ginseng saponin on the activity, phosphorylation, [$^{3}$H] ouabain binding and light scattering (disruption) of purified $Na^{+}$ ,$K^{+}$ -ATPase isolated from the outer medulla of sheep kidney were compared to those of gypsophila saponin, sodium dodecylsulfate (SDS), and Triton X-100 on the same parameters. $Na^{+}$ , $K^{+}$ -ATPase activity, phosphorylation, and [$^{3}H$] ouabain binding were inhibited by ginseng saponin (triol>total>diol), SDS, or Triton X-100, but increased by gypsophila saponin. Low doses of ginseng saponin (3.mu.g saponin/.mu.g protein) decreased phosphorylation sites and ouabain binding site concentration (Bmax) without any change of turnover number and affinity for ouabain binding which were decreased by high dose of ginseng saponin (over 10.mu.g saponin/.mu.g protein), SDS or Triton X-100. On the other hand, gypsophila saponin increased the affinity without any change of Bmax for ouabain binding. Inhibition of $Na^{+}$ ,$K^{+}$ -ATPase activity by ginseng saponin and SDS or Triton X-100 appeared before and after decrease in light scattering, respectively. These data suggest that ginseng saponins (total, diol, triol saponin) inhibit $Na^{+}$ , $K^{+}$ -ATPase activity by specific direct and general detergent action at low and high concentrations, respectively, and this inhibitory action of ginseng sapornin to $Na^{+}$ , $K^{+}$ -ATPase is not general action of all saponins.