• Applied Biological Chemistry
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• 제38권5호
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• pp.414-421
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• 1995

Lipoxygenase (LOX) 효소의 최적 활성 조건과 활성 억제등 LOX 활성 측정의 중요한 정보를 확립하기 위하여, 추출 Buffer의 영향, 기질, pH, 저장, 온도, NaCl, $CaCl_2$외 cations 및 antioxidants의 요소들이 LOX 활성에 미치는 영향을 조사하였다. 그리고 오이 tissue내의 LOX의 편재도 시험하였다. LOX에 대한 우수한 기질은 linolenic acid, linoleic acid, arachidonic acid 순서였다. 오이 껍질이나, mesocarp tissue내에 존재하는 LOX의 활성은 pH 5.5가 최적 조건이었으며, 섭씨 $40^{\circ}C$와 $50^{\circ}C$에서는 비교적 안정성을 보였다. LOX의 활성은 pH 5.0와 0.2M NaCl 조건을 같이 주었을때 opitimum 안정성을 보였다. LOX 활성은 $Mn^{2+},\;Cu^{2+}$ 또는 $Al^{3+}$와 같은 양이온에 의해서는 감소되었지만, 오히려 $Ca^{2+}$은 효소의 활성을 자극시켰다. 한편 butylated hydroxy anisole (BHA)와 propyl gallate의 농도가 증가할수록 LOX 활성은 감소되었다. 오이 껍질에서의 LOX의 활성은 다른 tissue에, locule, mesocarp, 비해 최고치를 보였다.

• 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.

• BMB Reports
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• 제46권5호
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• pp.237-243
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• 2013

Heart failure is one of the leading causes of sudden death in developed countries. While current therapies are mostly aimed at mitigating associated symptoms, novel therapies targeting the subcellular mechanisms underlying heart failure are emerging. Failing hearts are characterized by reduced contractile properties caused by impaired $Ca^{2+}$ cycling between the sarcoplasm and sarcoplasmic reticulum (SR). Sarcoplasmic/endoplasmic reticulum $Ca^{2+}$ ATPase 2a (SERCA2a) mediates $Ca^{2+}$ reuptake into the SR in cardiomyocytes. Of note, the expression level and/or activity of SERCA2a, translating to the quantity of SR $Ca^{2+}$ uptake, are significantly reduced in failing hearts. Normalization of the SERCA2a expression level by gene delivery has been shown to restore hampered cardiac functions and ameliorate associated symptoms in pre-clinical as well as clinical studies. SERCA2a activity can be regulated at multiple levels of a signaling cascade comprised of phospholamban, protein phosphatase 1, inhibitor-1, and $PKC{\alpha}$. SERCA2 activity is also regulated by post-translational modifications including SUMOylation and acetylation. In this review, we will highlight the molecular mechanisms underlying the regulation of SERCA2a activity and the potential therapeutic modalities for the treatment of heart failure.

• 한국산학기술학회논문지
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• 제15권11호
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• pp.6890-6897
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• 2014

본 전기로 환원슬래그는 팽창성 물질이 함유되어 불안정한 상태이므로 건설용 재료로 그대로 사용하기에는 다소 무리가 있다. 그러나 전기로 환원슬래그에는 $11CaO{\cdot}7Al_2O_3{\cdot}CaF_2$와 ${\beta}-C_2S$ (calcium aluminate compounds) 물질이 함유되어 있으므로 적절한 성분 보조제를 혼합하여 사용한다면 콘크리트용 혼화재료로서 사용이 가능할 것으로 판단된다. 이에 본 연구에서는 콘크리트 혼화재료서 전기로환원슬래그 미분말의 사용 가능성을 검토하고자 한다. 전기로 환원슬래그 미분말의 치환율별 모르타르 강도특성과 응결특성을 검토 결과, 최적 치환율은 30% 수준으로 나타났다. 최적 치환율에 석고 첨가에 따른 응결시간 및 강도특성을 파악한 결과, 전기로환원슬래그 미분말 치환율 30%인 배합에 이수석고를 8% 혼입하였을 경우 압축강도 발현 성상 및 응결특성이 OPC와 유사한 결과를 나타내었다.

• 대한약리학회지
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• 제2권2호
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• pp.35-42
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• 1966

The author investigated the effect of $Mg^#$, $Ca^#$, $Na^+$, $K^+$ and creatine phosphate on the ATPase activity of microsomal fraction isolated from rabbit uterus and obtained the following results : 1) The uterine microsomal fraction contained the $Na^+-$ and $K^+-$ activated ATPase in the presence of $Mg^#$. The ATPase activity increased with protein content in the fraction. 2) The maximum ATPase activity was obtained at $Na^+$ and $K^+$ concentraction of 100 mM respectively. 3) In the absence of $Mg^#$, the ATPase was not activated by $Na^+$ and $K^+$, but inhibited. 4) Car stimulated the $Na^+-$ and $K^+-$ activated ATPase in the presence of $Mg^#$. However, in the absence of $Mg^#$, the ATPase was not activated by $Ca^#$. 5) The $K^+-$ activated ATPase activity was greater than the $Na^+-activated$ ATPase under all conditions. 6) The $Na^+-$ and $K^+$ activated ATPase activity was increased by addition of creatine phosphokinase and creatine phosphate to the reaction mixture.

• 운동과학
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• 제26권3호
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• pp.229-237
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• 2017

PURPOSE: This study was to investigate the Glycolysis mediated sarcoplasmic reticulum (SR) $Ca^{2+}$ signal regulates mitochondria $Ca^{2+}$ during skeletal muscle contraction by using glycolysis inhibitor. METHODS: To examine the effect of Glycolysis inhibitor on SR and mitochondria $Ca^{2+}$ content, we used skeletal muscle fiber from gastrocnemius muscle. 2-deoxy glucose and 3-bromo pyruvate used as glycolysis inhibitor, it applied to electrically stimulated muscle contraction experiment. Intracellular $Ca^{2+}$ content, SR, mitochondria $Ca^{2+}$ level and mitochondria membrane potential (MMP) was detected by confocal microscope. Mitochondrial energy metabolism related enzyme, citric acid synthase activity also examined for mitochondrial function during the muscle contraction. RESULTS: Treatment of 2-DG and 3BP decreased the muscle contraction induced SR $Ca^{2+}$ increase however the mitochondria $Ca^{2+}$ level was increased by treatment of inhibitors and showed and overloading as compared with the control group. Glycolysis inhibitor and thapsigargin treatment showed a significant decrease in MPP of skeletal muscle cells compared to the control group. CS activity significantly decreased after pretreatment of glycolysis inhibitor during skeletal muscle contraction. These results suggest that regulation of mitochondrial $Ca^{2+}$ levels by glycolysis is an important factor in mitochondrial energy production during skeletal muscle contraction CONCLUSIONS: These results suggest that mitochondria $Ca^{2+}$ level can be regulated by SR $Ca^{2+}$ level and glycolytic regulation of intraocular $Ca^{2+}$ signal play pivotal role in regulation of mitochondria energy metabolism during the muscle contraction.

• 분석과학
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• 제25권5호
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• pp.333-338
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• 2012

IQGAPs는 세포 내에서 암세포화, 세포이동, 세포분열과 같은 다양한 기능을 수행하고 있으며, 대표적인 calmodulin (CaM) 결합 단백질로, human의 경우 3 개의 isoform이 알려지고 있다. IQGAPs는 각각 네 개의 IQ 부위를 가지고 있으며, 이들이 CaM과의 결합에 관여한다고 보고되고 있으나, 현재까지 IQGAP1에 비해 IQGAP3에서는 각각의 IQ 부위가 가지는 CaM 결합 특성에 대해선 연구가 미비한 실정이다. 따라서, 본 연구에서는 IQGAP3 내의 IQ 부위들과 CaM과의 결합성을 연구하였다. 이러한 연구를 수행한 결과, 네 개의 IQ부위가 의미 없는 CaM 결합성을 가지는 IQGAP1과는 다르게, IQGAP3는 IQ2와 IQ3가 $Ca^{2+}$-비의존성 CaM 결합을 보이고, IQ1과 IQ4는 결합성이 없음을 알 수 있었다. 또한, IQGAP1에서 새롭게 알려진 $Ca^{2+}$-의존성 CaM 결합 부위인 IQ(3.5-4.4) 부위가, IQGAP3에서도 잘 보존되어 있음을 알 수 있었다. 본 연구를 통해 IQGAP3의 IQ부위는 IQGAP1와는 다른 CaM 결합성이 있음을 알게 되었다. 이러한 결과는 각각의 IQGAP isoform들이 각기 다른 CaM 결합성으로 세포 내에서 다른 생리작용을 수행할 가능이 있음을 제시한다.

• Toxicological Research
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• 제17권
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• pp.127-133
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• 2001

Together with cytochrome P450 (CYP), flavin-containing monooxygenase (FMO) present in liver microsomes oxidizes various endogenous and exogenous chemicals. In an effort to determine the human FMO activity, we have developed two non-invasive urine analysis methods using caffeine (CA) and ranitidine (RA) as the probe compounds. As the production of theobromine (TB) and ranitidine N-oxide (RANO) from CA and RA is catalyzed primarily by the hepatic FMO, we have assigned the urinary molar ratios of TB/CA and RA/RANO as the in vivo FMO activity. In 200 age-matched Korean volunteers, the obtained TB/CA ratio ranged from 0.4 to 15.2 (38-fold difference) and the RA/RANO ratio from 5.7 to 27.2 (4.8-fold). The FMO activity of 20's, determined by caffeine metabolism, was the highest (2.5$\pm$l.9) and those of 30's, 40's, 50's, 60's and 70's were 40%, 50%, 24%, 39% and 36% of the 20's, respectively. Intake of grapefruit juice, known to contain flavonoids, inhibited the in vivo FMO (TB/CA) activity by 79%. Addition of the flavonoids like naringin, quercitrin and kaempferol, present in grapefruit juice, to the in vitro microso-mal FMO assay, thiobenzamide S-oxidation, produced 75%, 70% and 60% inhibition, respectively. Obtained Ki values of quercitrin, kaempferol and naringin on the in vitro FMO activity were 6.2, 12.0 and 13.9 $\mu\textrm{M}$, respectively. This suggested that the dose of drug should need to be adjusted to suit the individual FMO activities when the drugs metabolized by FMO are given to patients. As the intake of grapefruit juice has been identified to inhibit the FMO as well as CYP3A4 and lA2 activities, patients taking drugs metabolized by these enzymes should not drink grapefruit juice as the carrier.

• Natural Product Sciences
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• 제10권3호
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• pp.114-118
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• 2004

It is reported that $(1R,9S)-{\beta}-Hydrastine$ hydrochloride (BHSH) decreased the intracellular dopamine content by inhibiting tyrosine hydroxylase (TH) activity in PC12 cells. In this study, the inhibitory mechanisms on TH activity by BHSH in PC12 cells were investigated. BHSH treatment caused a reduction of TH activity and TH mRNA level in a dose-dependent manner. After the treatment of $20\;{\mu}M$ BHSH, TH activity and TH mRNA content were reduced at 15 min, reached the minimal levels at 6-24 h, and then recovered gradually to the control level. BHSH at $10-50\;{\mu}M$ caused a decrease in the basal intracellular cyclic AMP levels at 10 min in a concentration-dependent manner. In addition, BHSH at $20-100\;{\mu}M$ decreased the basal intracellular $Ca^{2+}$ concentration $([Ca^{2+}]_i)$ immediately in a dose-dependent manner. BHSH also inhibited the 56 mM $K^+ $ depolarization-induced elevation in $[Ca^{2+}]_i$, and blocked caffeine-activated store-operated $Ca^{2+}$ entry in PC12 cells. These data suggest that BHSH inhibits TH activity and TH gene expression, in part, through reducing cyclic AMP content and basal $[Ca^{2+}]_i$ in PC12 cells.

• BMB Reports
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• 제28권5호
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• pp.387-391
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• 1995

In order to investigate whether phospholipase C (PLC) activity in oat celIs is regulated by Gprotein, we have characterized PLC in plasma membranes of oat tissues. To identify the purified plasma membrane, $K^+$-stimulated, $Mg^{2+}$-dependent ATPase activity was measured. The activity of ATPase was shown to be proportional to the concentration of membrane protein. To examine the PLC activity regulated by G-protein, we used the inside-out and outside-out plasma membrane mixture isolated from the oat cells. The plasma membrane mixture showed higher PLC activity than the one of the outside-out plasma membrane. This suggests that PLC activity is located at the cytoplasmic surface of plasma membrane. PLC activity in plasma membrane mixture was dependent on $Ca^{2+}$ with maximum activity at 100 ${\mu}m$ $Ca^{2+}$ and it was inhibited by 1 mM EGTA. Using Sep-pak $Accell^{TM}$ Plus QMA chromatography, we found that inositol 1,4,5-trisphosphate ($IP_3$) was produced in the presence of 10 ${\mu}m$ $Ca^{2+}$. The PLC activity in the membrane was enhanced by an activator of G-protein ($GTP{\gamma}S$) and not by an inhibitor ($GDP{\beta}S$). This indicates that a G-protein is involved in the activation of PLC in the plasma membrane of oat cells.