• Fisheries and Aquatic Sciences
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• 제9권4호
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• pp.146-152
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• 2006

Systemic infections of maricultured fishes by Megalocytivirus species have occurred over a broad area in South Korea, causing extensive economic loss. We developed digoxigenin-labeled nucleic acid probes against the 230-bp ATPase and 311-bp major capsid protein (MCP) of rock bream Oplegnathus fasciatus iridovirus (RBIV) using polymerase chain reaction, and an in situ hybridization (ISH) method to detect Megalocytivirus in formalin-fixed tissues of mariculture species (rock bream, sea bass, and olive flounder). ISH-positive cells were abundant in the hematopoietic and connective tissues of various organs, while brain tissue showed little or no signal. The ISH procedure can become an important diagnostic tool in complement with histopathological methods, and advances epidemiological studies on the origin and distribution of Megalocytivirus in mariculture.

• 한국환경보건학회지
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• 제19권4호
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• pp.83-89
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• 1993

To evaluate an effect of methanethiol on a cause of erythrocyte membrane damage in rats, methanethiol was given at 11.25 rag/100 g body weight, and after 4 hr, the animals were sacrifled, the activities of Na$^+$/K$^+$ ATPase, protein contents in partial purified erythrocyte membrane and erythrocyte indices were determined Concomitantly, in vitro, effect of methanethiol on the erythrocyte fragility, Na$^+$/K$^+$ ATPase activity and its kinetics in various concentration of substrate from the preincubated erythrocyte membrane with methanethiol were demonstrated. The spleen weight per body weight (%) and MCV of erythrocyte in methanethiol-treated rats were more increased than those in the control group. The Na$^+$/K$^+$ ATPase activities in erythrocyte membrane were more decreased in methanethiol-treated rats than those in the control group. The apply of 0.05 ng rat whole blood to the 0.24 mg/ng of methanethiol solution in isotonic condition showed the complete hemolysis. The Na$^+$/K$^+$ ATPase activity in preincubated erythrocyte membrane with methanethiol at 37$\circ$C showed the dual effect and the K$_m$ value of Na$^+$/K$^+$ ATPase was higher in the preincubated erythrocyte membrane with methanethiol than that in the preincubated erythrocyte membrane omitted the methanethiol. These results suggest that the methanethiol may induce the damage of rat's erythrocyte membrane due to a change in substrate binding affinity of Na$^+$/K$^+$ ATPase.

• Biomolecules & Therapeutics
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• 제14권3호
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• pp.143-147
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• 2006

In the mouse leukemic monocyte cell line RAW 264.7, the vacuolar-type $(H^+)$-ATPase (V-ATPase) inhibitor bafilomycin $A_1$ at 10 and 100 nM decreased cell growth and survival as determined by 3-(4,5-dimethyl(thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in a concentration-dependent manner. At such concentrations, bafilomycin $A_1$ induced nitric oxide (NO) production through the expression of inducible nitric oxide synthase (iNOS). The bafilomycin $A_1$-induced NO production was inhibited by the NOS inhibitor $N^G$-monomethyl-L-arginine acetate (L-NMMA). Our findings suggest that the V-ATPase inhibitor bafilomycin $A_1$ induces NO production through the expression of iNOS protein.

• Journal of Acupuncture Research
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• 제18권2호
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• pp.150-160
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• 2001

Objectives ; This study was undertaken to determine whether Carthami-Flos aquacapuncture (CFA) exerts protective effect against oxidant-induced inhibition of $Na^+-K^+$-ATPase activity in cerebral synaptosomes. Methods and Results ; The enzyme activity was dependent on incubation time and enzyme protein concentrations. An oxidant t-butylhydroperoxide (tBHP) at 1 mM concentration caused a significant inhibition of $Na^+-K^+$-ATPase activity, which was prevented by addition of 0.01% CFA. tBHP inhibition and CFA protection were independent on incubation time or enzyme protein concentrations. The enzyme activity was increased by ATP in a dose dependent manner. Effects of tBHP and CFA were not affected by ATP cocentrations. tBHP (1 mM) produced a significant increase in lipid peroxidation in cerebral synaptosomes, which was prevented by 0.01% CFA. CFA decreased oxygen free radicals generated induced by the phorbol-ester in a dose-dependent manner in human neutrophil. Conclusions ; These results suggest that CFA exerts protective effect against tBHP-induced inhibition of $Na^+-K^+$-ATPase activity, which is due to by an antioxidant action resulting from a direct scavenging effect of oxygen free radicals in the cerebral synaptosomes.

• 동아시아식생활학회지
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• 제16권4호
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• pp.453-457
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• 2006

This study investigated the extractability, solubility, Mg$^{2+}$-, Ca$^{2+}$- and EDTA-ATPase activity of actomyosin prepared from leg and breast muscle of dog meat. The actomyosin extractability of breast muscle(2,100.6 mg/l00 g) was higher than that of leg muscle(500.8 mg/l00 g). The Mg$^{2+}$-ATPase activity of actomyosin had a high ionic strength of 0.02$\sim$0.05 M KCI and did not differ between leg and breast muscle. The Ca$^{2+}$-ATPase activity of actomyosin had a high ionic strength of 0.02$\sim$0.10 M KCI and leg muscle had a higher level of Ca$^{2+}$-ATPase activity than breast muscle did. The EDTA-ATPase activity was lower in low ionic strength and showed higher in high ionic strength, and increased sharply with increasing ionic strength up to 0.3 M KCI. The solubility of actomyosin did not differ between leg and breast muscle, and the solubility started and ended at KCI concentrations of 0.35 M and 0.4 M, respectively.

• 대한약리학회지
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• 제30권1호
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• pp.117-124
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• 1994

Phospholamban은 심근 근장그물 $Ca^{2+}-ATPase$ 조절단백이다. 조절작용기전은 탈인산화된 phospholamban에 의해 $Ca^{2+}-ATPase$가 억제됨으로 나타나며, 이 phospholamban이 인산화됨으로 $Ca^{2+}-ATPase$에 대한 억제가 반전됨을 보인다. 최근에 phospholamban의 cytoplasmic domain만으로 $Ca^{2+}-ATPase$를 억제하기에는 불충분하다는 보고가 있어 본 실험을 계획하였다. $Ca^{2+}-ATPase$의 활성을 억제하는 phospholamban domain을 밝히기 위하여 합성한 phospholamban 펩타이드(아미노산 1-25)의 $Ca^{2+}$ uptake에 대한 효과를 살펴보았다. 골격근 근장그물에서 $Ca^{2+}-ATPase$를 분리한 후 phosphatidylcholine이나 phosphatidylcholine과 phosphatidylserine을 포함한 liposome에 재조합시켰다. Phospholamban 펩타이드는 phosphatidylcholine을 이용하여 재조합된 vesicles의 초기 $Ca^{2+}$ uptake rate를 억제하고, cAMP 의존성 protein kinase의 catalytic subunit로 인산화시킨 phospholamban 펩타이드는 이 억제를 반전시킴을 보여 주었다. Phosphatidylcholine과 phosphatidylserine을 포함한 제조합 vesicles에서도 같은 양상을 보였다. 이상의 결과로 미루어 볼 때 인산화 sites를 포함하고 있는 phospholamban의 cytoplasmic domain은 그 자체만으로도 근장그물 칼슘펌프를 억제하기에 충분하다고 결론지을 수 있다.

• 한국미생물·생명공학회지
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• 제33권4호
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• pp.255-259
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• 2005

본 실험에서는 HRF의 조절단백질을 알아보기 위해 HRF를 bait로 한 yeast two hybrid assay를 실행한 결과 해당과정에 관여하는 TPI(triosephosphate isomerase)라는 효소를 발견하였으며, 가장 많이 중복되어 있었다. In vitro에서 HRF는 TPI의 C말단 잔기 부근(아미노산 156-249)이 상호작용에 주로 관여하는 부위임을 알 수 있었다. 또한, HeLa 세포에서 immunoprecipitation을 이용하여 HRF와 TPI의 상호작용이 실제 in vivo에서도 일어나는 현상이라는 것을 밝혔다. 결과적으로 HRF와 TPI 상호작용은 세포내 일정량이 존재하며 여러가지 신호전달에 의해 동시에 Na,K-ATPase와도 상호작용하는 것으로 생각된다.

• BMB Reports
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• 제33권4호
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• pp.289-293
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• 2000

Annexin I is a 37 kDa member of the annexin family of calcium-dependent phospholipid binding proteins. Annexin I plays regulatory roles in various cellular processes including cell proliferation and differentiation. Recently we found that annexin I is a heat shock protein (HSP) and displays a chaperone-like function. In this paper we investigated the function of annexin I as an ATPase using 1 to 32 amino acids deleted annexin I (${\Delta}-annexin$ I). ${\Delta}-Annexin$ I hydrolyzed ATP as determined by thin layer chromatography. The ability of ATP hydrolysis was inhibited by ADP, GTP and GDP, but not by the AMP, GMP and cAMP. In view of the ATP hydrolyzing function of HSP, the results support the function of annexin I as a HSP.

• 한국균학회지
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• 제19권3호
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• pp.214-219
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• 1991

1. 표고버섯, L. edodes 중의 adenosine-5'-triphosphatase를 황산암모늄 30% 포화로 분별침전 및 Sephadex G-200 겔 여과로 정제하였다. 2. 이 버섯 중에는 3종류의 단백질 분획과 adenosine-5'-triphosphate 기질에 대한 두 종류의 활성분획 I, II, III 및 IV를 얻었다. 3. 활성분획 II를 정제하여 얻은 이 효소의 최적 pH 및 최적 온도는 각각 7.6 및 $58^{\circ}C$였고, 열 안정성은 $20-40^{\circ}C$에서 30분 동안 안정하였다. 이 효소의 Km값은 1.81mM이었다.

• 한국동물학회지
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• 제23권2호
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• pp.61-68
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• 1980

발정주기에 EK라서 생쥐자궁의 alkaline phosphatase와 transport ATPases의 활성변화를 알아보기 위하여 정량적으로 분석하였다. 발정주기의 각 시기에 있어서 이 효소활성들의 비율은 대체로 그 양상이 서로 비슷하나, 발정기의 $K^+$-dependent와 $Na^+, K^+$-activated ATPases를 제외한 다른 효소들의 활성은 다른 어떤 시기보다도 유의하게 (p<0.025) 높았다. 즉, $K^+$-dependent와 $Na^+, K^+$-activated ATPases의 활성은 발정간기에서 발정기에 이르는 동안 무시할 정도이고, 다만 발정후기에 약간의 활성(0.04$\\sim$0.05 $\\mu$M/mg protein/hr), 총활성의 6$\\sim$7%)이 나타났다. 한편, 발정기에서 $Mg^++$-dependent phosphatase, transport ATPase와 alkaline phosphatase의 활성들은 급속히 현저하게 증가하였으며 각각 0.69(35%), 0.42(21%), 1.58(79%)였다. Alkaline phosphatase는 전 발정주기를 통해 0.60$\\sim$1.58(79$\\sim$90%)의 활성을 보여 그 주종을 이루었다. Alkaline phosphatase의 활성중에는 $Mg^++$-dependent의 것이 총활성의 12$\\sim$16%로 추정되었다. 그러므로 $K^+$-dependent와 $Na^+$-activated ATPases는 발정기 때에 자궁액의 누적을 조절하는 요인이 아니고 발정후기에는 자궁상피 속으로 내액을 재흡수하는 요인인 것으로 짐작되며, EH한 $Mg^++$-dependent phosphatase, transport ATPase 그리고 alkaline phosphatase는 생쥐의 자궁 상피세포에서부터 내액을 분지하는 데에 밀접히 관련되어 있는 것으로 사려된다.