• 한국균학회지
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• 제21권3호
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• pp.165-171
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• 1993

표고버섯 중의 광감응성 mitochondrial ATP synthase는 0.1 mM $Fe^{2+}$ 단독 이온에 의하여 그 활성이 대조구에 비해 102%, 증가되었으며, 반면 $Fe^{3+}$ 및 $Mg^{2+}$ 이온은 효소의 활성을 억제시켰다. 0.5 mM $Mg^{2+}$ 존재하에서 0.1 mM $Fe^{2+}$ 이온에 의한 이 효소의 활성은 32% 증가되었으며 0.5 mM $Mg^{2+}$ 존재하에서 $Fe^{3+}$ 이온효과는 단독 $Fe^{3+}$ 이온의 효과와 유사한 경향으로 효소의 활성을 저해하였다. 0.5 mM $Mg^{2+}$과 0.1 mM, 0.5 mM 및 1.0 mM $Fe^{3+}$ 이온의 공존하에서$Fe^{2+}$ 이온에 의한 효소의 활성은 모두 억제되었으며, 특히 0.5 mM $Mg^{2+}$과 0.1 mM $Fe^{3+}$ 이온의 공존하에서 5.0 mM $Fe^{2+}$ 이온에 의하여 53%의 억제현상을 나타내었다. 따라서 표고버섯 중의 광감응성 mitochondrial ATP synthase의 활성은 $Fe^{2+}$ 이온에 의하여 특이적으로 크게 증가되며, 이 효소에 대한 $Fe^{2+}$ 이온의 활성화 효과가 $Mg^{2+}$ 이온에 의하여 크게 영항을 받지 않으나, $Fe^{3+}$ 이온의 공존하에서는 억제됨을 알았다. 활성화 금속이온인 $Fe^{2+}$ 존재하에서 이 효소의 최적 pH는 7.6이며, 최적 온도는 $63^{\circ}C$이었다. 또한 이 효소는 금속 chelating agent인 EDTA에 의하여 효소의 활성이 상실됨으로써 metalloenzyme의 가능성을 제시하였다.

• 약학회지
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• 제30권1호
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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.

• 분석과학
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• 제5권4호
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• pp.477-484
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• 1992

형질 전환된 사철느타리버섯으로부터 초원심분리 및 설탕밀도 기울기법으로 44% 층에서 미토콘드리아를 분리정제하였다. ATPase와 ATP synthase의 최적 활성조건은 각각 pH 7.4, $60^{\circ}C$ 및 pH 7.5, $57^{\circ}C$였고, km값은 11.6mM과 8.4mM였다. ATPase는 기질농도 5~6mM의 ATP에서, ATP synthase는 5~10mM ADP 농도에서 활성이 높으며, 그 이상의 농도에서는 기질 저해를 받았다. ATPase/ATP synthase 모두 $Mg^{2+}$ 의존성 효소로 $G_{418}$으로 비경쟁적인 저해를 받았다. 효소의 아미노산 분석결과 hydrophobic 아미노산 잔기는 50.5%, small 아미노산 잔기는 56.1%, hydrogen bonding 아미노산 잔기는 43.7%, helix breaking 아미노산 잔기는 55.2%였다. 인지질을 분석한 결과 phosphatidyl glycerol, phosphatidyl choline 및 phosphatidyl ethanolamine으로 구성되었고 phosphatidyl serine과 phosphatidyl inositol은 전혀 없었다. 포화 지방산은 palmitate(51.31%)와 stearate(18.32%)의 함량이 많았고, 불포화 지방산($C_{18:1}$, $C_{18:2}$ 및 $C_{16:1}$)의 함량도 많았다.

• 한국수산과학회지
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• 제17권4호
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• pp.271-279
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• 1984

이스라엘 잉어의 근육 actomyosin의 열안정성과 그 첨가제에 의한 영향을 밝히기 위하여 배육골격근에서 추출한 actomyosin을 시료로 하여 온도의 변화가 actomyosin의 안정성에 미치는 영향을 ATP-감도, 유리 SH기 및 Ca-ATPase 활성도 등을 측정하여 분석하였다. 그리고 sucrose, sorbitol, Na-glutamate 및 L-cysteine등 첨가제의 영향에 대하여는 Ca-ATPase 활성도의 변화를 측정하여 단백질 변성속도당수($K_D$), 단백질변성보호효과(${\Delta}E/M$) 그밖에 열력학적 제상수를 계산 비교하였다. 실험결과를 요약하면 다음과 같다. 1. 배육골격근에서 추출한 이스라엘 잉어 actomyosin은 단백질농도 $4.12{\sim}4.68mg/ml$, 핵산의 함량 $2.63{\sim}2.9%$, actin과 myosin의 결합비 $1:2.20{\sim}2.63$, 지질의 함량 $4.33{\sim}5.26\%$, ATP-감도 109.78, Ca-ATPase 활성 $0.159{\sim}0.201\;{\mu}M-Pi/min/mg-protein$, 유리 SH 기 함량 $3.3{\sim}3.4M/10^5g-protein$ 이었다. 2. Ca-ATPase활성 및 ATP-감도는 온도가 상승 함에 따라 1차반응적으로 감소하였고, 유리 SH기는 $60{\circ}C$ 까지는 증가하다가 그 이후는 급격히 하강하였다. 3. 가열온도상승에 따른 Ca-ATPase 활성의 반감 시간은 $12^{\circ}C$ 일때 280분, $20^{\circ}C$ 일 때 125분, $30^{\circ}C$일때 55분, $40^{\circ}C$ 일때 13분이었으며 $20^{\circ}C$에서 활성화에너지는 5,395 cal/mole 활성화엔탈피는 4,814cal/mole, 활성화엔트로피는 -40.42 e.u, 자유에너지 값은 17,626cal/mole이었다. 4. 당 및 아미노산중에서 가열에 대하여 변성보호효과가 높은 것은 $3\%$ sorbitol이었으며, $8\%$ Na-glutamate, $1\%$ sucrose, $1\%$ L-cysteine의 순으로 낮아졌다. 5. 저온저장시 actomyosin이 가장 안정한 온도는 $-30^{\circ}C$이었으며, $0^{\circ}C,\;-20^{\circ}C$ 순으로 불안정하였다. 또한 $-20^{\circ}C$ 일 때의 첨가제에 의한 냉동변성보호효과는 $8\%$ Na-glutamate가 가장 좋았고 $3\%$ sorbitol, $1\%$ sucrose, $1\%$ L-cysteine의 순으로 효과가 떨어졌다.

• Archives of Pharmacal Research
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• 제19권5호
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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.

• The Korean Journal of Physiology
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• 제10권1호
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• pp.35-40
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• 1976

In an attempt to examine the effect of ginseng on sodium transport across the biological membrane, we have studied effects of ginseng alcohol extract on the short-circuit current(SCC) and the $Na^+-K^+$-activated ATPase activity in isolated frog skin preparations. 1. Ginseng alcohol extract applied to the mucosal surface of the frog skin significantly increased SCC at low concentration($1{\sim}10mg%$) but decreased SCC at higher concentration($50{\sim}250mg%$). 2. Similarly, when the drug was added to the serosal bathing medium, the SCC was stimulated at low doses($5{\sim}25mg%$) and inhibibited at high doses($50{\sim}250mg%$). 3. $Na^+-K^+$-activated ATPase activity of the frog skin epidermal homogenate was significantly inhibited by ginseng alcohol extract, the effect being proportional to the concentration of the drug in the incubation mixture. These results may suggest that a low dose of ginseng alcohol extrat enhances the transepithelial sodium transport probably by increasing the permeability of outer membrane of the transporting cell to sodium ion, whereas a high dose of drug reduces the sodium transport primarly by inhibiting $Na^+-K^+$ ATPase mediated active transport step.

• 한국식품영양과학회지
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• 제16권3호
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• pp.55-61
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• 1987

산란노계육(産卵老鷄肉)(Arbor Acres, 72주령(週齡))의 가슴근육과 다리근육을 냉장 및 동결저장하면서 pH, 단백질의 추출성, 근원섬유단백질의 ATPase 활성, 소편화, 냉동감량 및 드립량의 변화를 비교하였다. 가슴근육과 다리근육의 pH는 각각 냉장 1일 및 동결저장 1주에 가장 낮았다. 근원섬유단백질의 추출성은 냉장기간이 경과하면서 점차 증가하였으며, 동결저장에서는 1주에 가장 높은 수준을 보였다. 근원섬유단백질의 $Mg^{2+}-ATPase$활성은 냉장 1일 및 동결저장 1주에 각각 높게 나타났다. 근원섬유의 소편화 정도는 냉장 1일 및 동결저장 1주에 크게 변화하였으며 냉동감량과 드립량은 동결저장 기간이 경과하면서 점차 많아졌다. 냉장 및 동결저장 중 가슴근육은 다리근육에 비하여 pH가 낮았고, 근원섬유단백질의 추출성, $Mg^{2+}-ATPase$활성, 드립량 및 소편화 정도가 높았으나 저장기간 중 변화되는 양상은 비슷하였다.

• 대한약리학회지
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• 제19권1호
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• pp.101-106
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• 1983

Vanadium is widely distributed in animal tissues and it is supposed to be a regulator of Na-K-ATPase activity. The effect of sodium orthovanadate on Na-K-ATPase activity in rabbit kidney was measured in vitro and compared with that of ouabain. The influence of sodium orthovanadate on the renal function of rabbits was also investigated. 1) Na-K-ATPase activity was decreased by sodium orthovandate at the concentrations of $10^{-7},\;10^{-6},\;10^{-5}\;and\;10^{-4}\;M$ to 73.89, 36.49, 6.50 and 4.99% of the control activity respectively. 2) Na-K-ATPase activity was decreased by ouabain at the concentrations of $10^{-4},\;10^{-3}\;and\;10^{-2}\;M$ to 69.52, 22.84 and 3.88% of the control activity respectively. 3) Urine volume, urinary excretion of $Na^+,\;K^+\;and\;Cl^-$, clearances of inulin and p-amino-hoppuric acid were decreased until after 60 minutes following the administration of sodium orthovanadate 0.5 mg/kg intravenously $Na^+\;reasorption$ rate was not changed and mean arterial pressure was significantly elevated during 60 minutes after the administration of sodium orthovanadate.

• 대한한방내과학회지
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• 제15권1호
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• pp.80-98
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• 1994

According to the original documents, Sagungsan is considered as an effective drug for controlling the hypertensive epistaxis induced by tension of autonomic nerve and it's hyperfunction. The present experiment was designed to understand the effect of Sagungsan extract on the hemostatic action, intracranial pressure, blood pressure and cardiovascular system in experimental animals. And thus the bleeding time, prothrombin time, capillary dilation, blood pressure, Intracranial pressure, and enzymatic analysis of the ATPase activities were studied. The result obtained here were as followings: 1. Sagungsan water extract reduced the bleeding time in mouse, and prolonged the prothrombin time in rabbits. 2. The drug extract increased the tail volume by capillary dilation in rats. 3. The drug extract inhibited the increase of intracranial pressure and arterial blood pressure in rabbits. 4. At the early time, the increase of arterial blood pressure by the drug extract significantly inhibited by pretreated atropin and regitine in rabbits. 5. The drug extract relaxed the smooth muscle by stimulating the Mg2+-Ca2+-ATPase activities of gastric sarcoplasmic reticulum isolated from rabbit stomach. 6. The drug extract stimulated the heart contraction by inhibiting the $Mg^{2+}-Ca^{2+}-ATPase$ activities of cardiac sarcoplasmic reticulum isolated from rabbit heart. The inhibitory mechanism was reversible and noncompatitive. 7. The drug extract increased the hepatic blood volume by stimulating the hepatic total ATPase activities and hepatic metabolism. 8. The drug extract acted as a tranquilizer by inhibiting the neural Na+-K+-ATPase activity. According to the results, Sagungsan water extract dilated the capillaries, stimulated the heart beat, and thus increased the blood flow with decreasing the intracranial pressure and blood pressure. These effects stanches the epistaxis collectively.

• 대한수의학회지
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• 제23권1호
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• pp.9-16
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• 1983

The effects of ethanol on Na-K-ATPase activity were investigated with cat kidney homogenate. The results were summarized as follows: 1. Na-K-ATPase activity was inhibited with dose-dependent manner by ethanol of higher concentration than 1%, and showed an estimated $I_{50}$ (the inhibitor concentration to cause 50% inhibition) of 7.5%. 2. Hydrolysis of ATP was linear with the incubation time in the absence and presence of 8% ethanol, whereas it was different with preincubation time in the presence of 15% ethanol. 3. Inhibition of Na-K-ATPase activity by ethanol was not affected by increased enzyme concentration, and showed the reversibility of the inhibitory pattern. 4. Kinetic studies of cationic-substrate activation of Na-K-ATPase showed that ethanol had both properties of classical competitive inhibition for $Mg^{{+}{+}}$ or $K^+ and non-competitive inhibition for ATP or $Na^+$. 5. Arrhenius plot yield two break point at $21^{\circ}$ and $30^{\circ}C$ in the absence of ethanol, whereas showing only one break point at $18^{\circ}C$ in the presence of 8% ethanol. These results suggested that ethanol inhibited Na-K-ATPase activity reversible through a disturbance of microenvironment of lipids associated with the enzyme.