• 한국수산과학회지
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• 제30권3호
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• pp.349-354
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• 1997

6개월, 12개월 및 20개월간 사육한 넙치를 즉살하여 암수를 구분한 후 등육을 취하여 각각의 사육기간 별로 근원섬유의 ATPase 활성, 열안정성 및 각 단백질의 subunit조성을 실험, 검토하였다. 6개월 사육한 수컷의 근원 섬유의 ATPase 활성은 암컷에 비하여 높았으며, 특히 $Mg^{2+}\;(+Ca^{2+})$-와 $Ca^{2+}-ATPase$ 활성에서 현저한 차이를 나타내었다. 12개월간 사육한 넙치에 있어서도 6개월간 사육한 것과 비슷한 경향을 보였으며, 20개월간 사육한 넙치 역시 유사한 경향을 나타내어 주었다. 또한 사육기간에 따라서 근원섬유의 ATPase 활성이 차이를 나타내었는데, 특히 $Mg^{2+}\;(+Ca^{2+})-ATPase$ 활성이 성장이 활발한 6개월째 사육한 넙치에서 가장 높았으며, 그 다음으로 12개월과 20개월간 사육한 순으로 나타나 성장 속도와 근원섬유의 ATPase 활성간에 높은 상관관계가 있음을 암시하였다. 근원섬유의 열안정성에 있어서도 수컷이 암컷에 비하여 현저히 떨어지는 경향을 보였다.

• 한국동물학회지
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• 제37권4호
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• pp.531-537
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• 1994

Partial internal amino acid sequences of the 15S ATPase from chick skeletal muscle were determined and found to be identical to the corresponding regions of the Mg2+_ATPase from Xenopus laevis oocytes, that is a close homolog of N-ethvlmaleimide-sensitive factor (called NSF) in hamster and Sec18p in yeast, both of which are believed to plaN an essential role in vesicle fusion in secretory process. Thus, the 15S Arpase in chick skeletal muscle maw also belong to a protein family of the "vesicle fusion proteins". Unlike the Mg2'-Afpase with an isoelectric point (pl) of 5.5, however, the 15S Arpase was separated into four spots with pls of 4.9,6.4 and 6.9 upon analysis by twoiimensional gel electrophoresis. In addition, the anti-15S ATPase IgG was found to be capable of interacting with the 265 protease complex upon analysis by immunoprecipitation. Moreover, immunoblot analysis revealed that the anti-155 Arpase IgG recognizes three subunits, ko of which show the same mobilities as the 510-kDa subunit 4 and 48-kDa subunit 7 of the 26S protease complex that are known to contain a highly consented ATP-binding motif. These results surest that a common antigenic site, likely the consensus nucleotide-binding site, exists in the 15S ATPase and the 26S pretense complex and hence both the enzymes maw also be related ATPases.d ATPases.

• The Korean Journal of Physiology
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• 제29권2호
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• pp.269-277
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• 1995

Membrane vesicles were prepared by differential centrifugation from epithelial cells of porcine trachea. Total activity of microsomal ATPases was measured spectrophotometrically by a coupled enzyme assay. The steady-state activity of the enzyme was $329{\pm}10$ nmol/min mg protein. Thapsigargin, a specific antagonist of intracellular $Ca^{2+}-ATPase$, inhibited about 50% of the activity, leaving $178{\pm}18\;nmol/min .mg$ protein (n=6), indicating that the $Ca^{2+}-ATPase$ is one of the major microsomal ATPases. The microsomes used in this study appeared to be tight-sealed vesicles since they showed saturation in $^{45}Ca^{2+}$ uptake experiments. Inositol 1,4,5-trisphosphate $InsP_{3}, 4\;{\mu}M$, an agonist of $InsP_{3}$-sensitive $Ca^{2+}$ release channel ($InsP_{3}$, receptor), and Ca-ionophore A23187 $(10\;{\mu}M)$ induced $^{45}Ca^{2+}$ releases of 20% and 50% of stored $^{45}Ca^{2+}$, respectively. The addition of $(10\;{\mu}M\;InsP_{3}$ also increased the microsomal ATPase activity from $282{\pm}8$ nmol/min mg protein to $334{\pm}21$ nmol/min . mg protein in the intact vesicles. Similar increase in the activity was observed by making microsomes leaky (uncoupling) using the Ca-ionophore A23187. ;$InsP_{3}-induced$ effects were blocked by either thapsigargin or heparin suggesting that: 1) the $InsP_{3}-induced$ increase in ATPase activity is mediated by microsomal $Ca^{2+}-ATPase$, and 2) dissipation of $Ca^{2+}$ gradient across the microsomal membrane is responsible for the $InsP_{3}-induced$ effect. In order to test the dependence of the $Ca^{2+}-ATPase$ activity on the activity of $InsP_{3}-induced$ the activity of ATPases was monitored in various concentrations of free $Ca^{2+}$ using $EGTA-Ca^{2+}$ buffers. The $Ca^{2+}$-dependent biphasic change is the well-known character of $InsP_{3} receptor but not of microsomal $Ca^{2+}-ATPase$ in non-excitable cells; however, the activity of microsomal ATPase appeared biphasic and a maxim진 activity of $397{\pm}36nmol/min\;.mg$ protein was obtained in the solution containing 100 nM free $Ca^{2+}$. Below or above this concentration, the activity of ATPases was lower. These results strongly support a positive correlation of microsomal $Ca^{2+}-ATPase$ to the $InsP_{3}$ receptors in epithelial microsomes.

• 한국미생물·생명공학회지
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• 제8권2호
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• pp.113-117
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• 1980

식물성 식용유를 기본식이에 첨가하여 일정한 조건하에서 사육한 토끼 근육에서 근원섬유 단백질을 추출하여 ATPase활성과 이에 미치는 EDTA, $Ca^{2+}$, $Mg^{2+}$ 등의 여러가지 농도변화에 따른 한성도 저해 양상을 조사한 결과는 다음과 같다. 1. 근원섬유 단백질의 ATPase 환성은 KCI의 농도가 크면활성은 감소되고, 농도가 감소되면 황성은 증가되었으며 대조군보다 식용유를 급여한 군이 더 높았다. 2, EDTA의 농도변화에 따른 ATPase 학성은 0.2mM EDTA 이상부터 찬성방해작용이 현저히 나타났다. 3. $Ca^{2+}$, $Mg^{2+}$이 ATPase 활성에 미치는 영향을 보면 0.2mM $Ca^{2+}$ 이상의 농도에서부터, 1.0mM $Mg^{2+}$ 이상의 농도에서부터 ATPase 활성 방해작용이 뚜렷이 나타났다. 4. In vitro 소화율은 pepsin으로 처리하여 대조군이 71.66%, 들깨기름 급여군이 70.62%, 콩기름 급여군이 67.93%, 미강유 급여군이 86.79% 이었고, Trypsin으로 처리하면 대조군이 73.87%, 들깨기름 급여군이 77.93% 콩기름 급여군이 76.52%, 미강유 급여군이 90.22%를 보였다. 이는 pepsin에 의한 소화보다 Trypsin에 의한 소화력이 더 좋음을 나타내며 식물성 기름을 급여한 군의소화율이 더 좋음을 알 수 있었다.

• 생명과학회지
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• 제7권4호
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• pp.336-341
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• 1997

This study was conducted to investigate the effects of carcass grade on the hardness, myofibrillar fragmentations index, protein extractability and Mg-ATPase activity of myofibril and actomyosin obtained from 1, 2, 3 and D carcass grade)subgrade) in Korean native cow. Proximate component, hardness, chewiness, myofibril fragmentation index, protein extractability and Mg-ATPase activity if myofibril or actomyosin were not significantly different between 1st and 2nd carcass grade loin. The hardness and chewiness of 2nd carcass grade loin's were significantly lower than 3th grade loin's, but the myofibril fragmentation index, sarcoplasmic protein extractability and Mg-ATPase activity of myofibril were higher. The myofibrillar protein extractability and Mg-ATPase activity of actomyosin obtained from 3th carcase grade loin's were significantly higher than D grade loin's, but the hardness, chewiness and stroma protein extractability were lower. In conclusion, the degree of toughness in Korean native cow's loin was not significantly different between 1st and 2nd grade, but 3rd and D carcass grade were significantly higher, regardless of before and after aging.

• BMB Reports
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• 제29권1호
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• pp.22-26
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• 1996

Agents that activate or inhibit the $Ca^{2+}$ release channel in cardiac sarcoplasmic reticulum (SR) were tested for their abilities to affect the activity of the SR $Ca^{2+}$-ATPase. Vesicles of junctional SR (heavy SR, HSR) from terminal cisternae were prepared from porcine cardiac muscle by density gradient centrifugation. The steady-state activity of $Ca^{2+}$-ATPases in intact HSR vesicles was/$347{\pm}5\;nmol/min{\cdot}mg$ protein (${\pm}$ SD). When the HSR vesicles were made leaky, the activity was increased to $415{\pm}5\;nmol/min{\cdot}mg$ protein. This increase is probably due to the uncoupling of HSR vesicles. Caffeine (10 mM), an agonist of the SR $Ca^{2+}$ release channel, increased $Ca^{2+}$-ATPase activity in the intact HSR vesicle preparation to $394{\pm}30\;nmol/min{\cdot}mg$ protein. However, caffeine had no significant effect in the leaky vesicle preparation and in the purified $Ca^{2+}$-ATPase preparation. The effect of caffeine on SR $Ca^{2+}$-ATPase was investigated at various concentrations of $Ca^{2+}$. Caffeine increased the pump activity over the whole range of $Ca^{2+}$ concentrations, from $1\;{\mu}M$ to $250\;{\mu}M$, in the intact HSR vesicles. When the SR $Ca^{2+}$-ATPase was inhibited by thapsigargin, no caffeine effect was observed. These results imply that the caffeine effect requires the intact vesicles and that the increase in $Ca^{2+}$-ATPase activity is not due to a direct interaction of caffeine with the enzyme. We propose that the activity of SR $Ca^{2+}$-ATPase is linked indirectly to the activity of the $Ca^{2+}$ release channel (ryanodine receptor) and may depend upon the amount of $Ca^{2+}$ released by the channels.

• 한국균학회지
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• 제21권3호
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• pp.157-164
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• 1993

1. 표고버섯중 광감응성 mitochondrial $F_1-ATPase$는 $Fe^{3+},\;Fe^{2-}$ 및 $Mg^{2+}$ 이온에 의하여 각각 활성화 되었으나 5.0 mM $Fe^{3+}$ 이온에 의한 상대활성도는 대조구에 비하여 107% 증가시켰다. 2 $Mg^{2+}$ 존재하에서 $Fe^{3+}$ 및 $Fe^{2-}$ 각 이온 농도효과는 모두 효소의 활성을 증가시켰으나 0.1 mM $Mg^{2+}$과 5.0 mM $Fe^{3+}$ 이온의 공존하에서 170%를 증가시켜 $Mg^{2+}$ 이온에 의한 상승작용을 보였다. 3. 0.1 mM $Mg^{2+}$와 0.1 mM $Fe^{2+}$ 존재하에서 $Fe^{3+}$ 이온농도효과는 그 농도가 5.0 mM일 때 168%의 활성도 증가를 보여 $Fe^{2-}$ 이온공존효과는 없었다. 4. 이 효소는 $Mg^{2+}$ 및 $Fe^{3+}$ 이온에 의하여 활성화되는 특성을 가지고 있으며 활성금속이온 존재하에서 측정한 최적 pH 빛 온도는 각각 7.5 및 $66^{\circ}C$였다.

• 대한한의학회지
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• 제17권2호
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• pp.264-276
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• 1996

This study was carried out to evaluate the effect of Samhwasan on the Na-K-ATPase activity of heart muscle. The Na-K-ATPase activity was prepared from rabbit heart ventricles. Samhwasan markedly inhibited the Na- K - ATPase activity in a dose-dependent manner with an estimated $I_{50}$ of 0.56%. Hill coefficient was 1.70, indicating that the enzyme has more than one binding site for the Samhwasan. Inhibition of enzyme activity by Samhwasan increased as pretreatment time was prolonged. Inhibition by the drug was not affected by a change in enzyme protein concentration. Kinetic studies of substrate activation of the enzyme indicated classical noncompetitive inhibition, showing significant reduction in Vmax without a change in Km value. Inhibitory effect by Samhwasan was not altered by changes in concentration of $Mg^{2+}$, $Na^+$ or $K^+$, dithiothreitol. a sulfhydryl reducing reagent, did not protect the inhibition of Na-K-ATPase activity by Samhwasan combination of Samhwasan and ouabain showed a cumulative inhibition fashion. These results suggest that Samhwasan inhibits Na-K-ATPase activity of heart ventricles with an unique binding site different from that of ATP, $Mg^{2+}$, $Na^+$ or $K^+$ and ouabain.

• 대한약리학회지
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• 제2권1호
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• pp.31-40
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• 1966

The microsomal fraction is isolated from rabbit heart and skeletal muscle. The fraction is found to contain the $Na^+$-and $K^+$-activated ATPase. The maximal ATPase activity is obtained in $Na^+$ and $K^+$ concentration of 100 mM. Calcium itself stimulates the $Na^+$-and $K^+$-activated portion of ATPase in the presence of $Mg^{++}$. However, calcium does not stimulate ATPase in the absence of $Mg^{++}$.

• 한국동물학회지
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• 제10권2호
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• pp.1-8
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• 1967

토끼의 골격근 homogenate에서 23,000$\times$G, 60 분간의 원심분리와 얻은 근 microsome의 ATPase 활성, 근수축에 대한 이완작용, 및 Ca 의 흡수작용을 여러 가지 조건에서 측정하였다. ATPase 활성은 Ca++ Mg++ 양 이온의 존재에 의하여 활성화되며 , 5 mM Mg++ 의 존재하에서는 Ca++ 의 최적농도는 0.1mM이다. Oxalate의 존재하에서는 1 mM 의 Ca++ 이 최적농도이므로 oxalate의 작용은 불용성 Ca-oxalate의 작용은 불용성 Ca-oxalate를 microsome vesicle so 및 medium 내에 침전시켜 유리 Ca++ 농도를 저하시키는 것이라고 생각된다. Microsome의 이완작용은 조제후 120 시간까지 시간에 따라 감소되어 가나, 그이 ATPase 활성은 거의 변화가 없는 것으로 보아 Ca++ + Mg++ -의존성 ATPase 는 이완작용에는 직접 관련이 없는 것으로 해석된다. Oxalatedmlwhswo는 microsome의 Ca++ 흡수량을 현저히 증대시키며 동시에 흡수포화에 도달하는 시간을 지연시킨다. Oxalate의 이러한 효과도 Ca-oxalate의 형성에 기인하는 것으로 해석된다. Microsome 내에 축적되는 Ca 의 량은 ATP 농도가 커질수록 많아진다. 그러나 축적된 Ca 의 량과 ATP 농도사이에 화학정량론적 관계는 없는 것같다.