• 생명과학회지
• /
• 제12권6호
• /
• pp.812-820
• /
• 2002

이 연구는 주변의 김치들을 수거하여 젖산에 내성을 가진 Leuconostoc mesenteroides을 선택배지를 이용하여 분리하고, 지방산 조성을 분석하여 동정한 결과, Leuconostoc mesenteroides LM3균주로 동정했다. 이어서 그의 성장패턴을 관찰하고, 유도기와 지수성장기에서 젖산을 첨가한 결과 성장저해가 일어났음을 확인했으며, 젖산의 최종농도가 0.4%이상인 배지에서는 성장속도가 느려짐을 알았다. 그리고 정지기시작단계에서 고농도의 젖산을 단시간 처리하였을 때, 대조균주가 생존하지 못하는 4%농도에서도 Leuconostoc mesenteroides LM3군주가 생존하는 것을 확인하여 산에 대한 내성이 뛰어남을 알 수 있었다. 이에 trehalose 농도의 변화, 그리고 trehalase와 ATPase 활성을 측정하였고, 이를 통해 젖산에 의한 세포내의 변화를 관찰하였다. 그 결과, 각 성장시기별로 0.6% 젖산을 첨가했을 때 Leuconostoc mesenteroides LM3균주가 trehalose 축적률이 높아졌으나 trehalase활성엔 큰 변화가 없었다. ATPase 활성은 감소하는 경향을 보였으나 시기별 활성은 일정하였고, 세포내 pH를 측정한 결과 Leuconostoc mesenteroides LM3균주는 pH 6으로 일정하게 유지되었다.

• 한국식품영양과학회지
• /
• 제24권6호
• /
• pp.867-873
• /
• 1995

만성적인 알코올 섭취가 미토콘드리아의 지질조성 및 ATPase 활성도에 미치는 영향을 조사하고 아울러 간의 발암물질인 2-AAF를 토여하여 간 미토콘드리아의 기능에 미치는 상호 효과를 조사하기 위하여 120~125g 숫쥐에게 열량의 35%를 알코올로 공급하여 6주간 사육하였으며 알코올 섭취 4주 후에 2-AAF를 3일 간격으로 2회 복강투여하여 실험한 결과를 요약하면 다음과 같다. 1. 체중 증가, 식이 섭취량 및 식이효율에 대한 알코올의 효과는 유의적인 차이를 보였으나, 2-AAF에 의한 유의적인 효과는 관찰할 수 없었다. 알코올 섭취군이 대조군 보다 체중은 유의적으로 감소하였다. 2. 미토콘드리아 ATPase 활성도는 알코올 섭취에 의해 유의적으로 감소되었고, 2-AAF 투여에 의해서는 유의적이지 않으나 감소되는 경향을 보였다. 따라서 대조군(C)에 비하여 알코올 섭취와 2-AAF투여군(EA)에서 ATPase 활성도가 29.3%의 감소를 나타냈다. 3. 간 미토콘드리아 콜레스테롤 함량은 알코올 섭취에 의해 유의적으로 증가하였으며 미토콘드리아 총 인지질량은 알코올과 2-AAF에 의한 상호작용에 의해 알코올 투여군에서만 2-AAF 투여시 유의적인 증가 효과를 보여주었다. C/PL 비율은 알코올 섭취와 2-AAF 투여 보두에서 유의적인 효과는 보이지 않았으나 군간 비교시 알코올군(E)이 대조군(C) 보다 유의적으로 증가하였다. 4. 간 미토콘드리아의 CL량은 알코올군에서 감소하는 경향을 보였고 PS와 PI량은 차이가 없었다. 알코올 섭취로 PC량은 유의적으로 증가하였으나 PE량은 차이가 없엇으며, 2-AAF 투여시 PC량은 유의적인 감소를 보였고 PE량은 유의적으로 증가하였다. PE/PC 비율은 알코올 투여에 의해 유의성은 없었으나 감소경향을 나타냈으며, 2-AAF 투여에 의해서는 유의적인 증가 효과가 나타났다. 5. 알코올 섭취시 미토콘드리아의 지방산 조성은 palmitic acid가 유의적으로 감소하였고, stearic acid는 유의적으로 증가함을 보였다. UFA/SFA와 UI는 알코올군에서 유의적이지는 않으나 증가되는 경향을 보였다. 2-AAF 투여에 의해서도 linoleic acid는 감소되고 arachidonic acid는 증가되는 경향을 보여 UI값이 증가되는 경향이 있었다. 따라서 본 실험에서 나타난 ATPase 호라성도 변화는 비토콘드리아의 콜레스테롤변화, C/PL 비율, 인지질 조성이나 지방산 조성의 변화 등 막지질 조성의 변화에 기인하는 것으로 생각된다.

• Animal cells and systems
• /
• 제14권1호
• /
• pp.1-10
• /
• 2010

Previously we showed that CHIP, a co-chaperone of Hsp70 and E3 ubiquitin ligase, can promote the degradation of mutant SOD1 linked to familial amyotrophic lateral sclerosis (fALS) via a mechanism not involving SOD1 ubiquitylation. Here we present evidence that CHIP functions in the interaction of mutant SOD1 with 26S proteasomes. Bag-1, a coupling factor between molecular chaperones and the proteasomes, formed a complex with SOD1 in an hsp70-dependent manner but had no direct effect on the degradation of mutant SOD1. Instead, Bag-1 stimulated interaction between CHIP and the proteasome-associated protein VCP (p97), which do not associate normally. Over-expressed CHIP interfered with the association between mutant SOD1 and VCP. Conversely, the binding of CHIP to mutant SOD1 was inhibited by VCP, implying that the chaperone complex and proteolytic machinery are competing for the common substrates. Finally we observed that mutant SOD1 strongly associated with the 19S complex of proteasomes and CHIP over-expression specifically reduced the interaction between S6/S6' ATPase subunits and mutant SOD1. These results suggest that CHIP, together with ubiquitin-binding proteins such as Bag-1 and VCP, promotes the degradation of mutant SOD1 by facilitating its translocation from ATPase subunits of 19S complex to the 20S core particle.

• The Korean Journal of Physiology
• /
• 제21권1호
• /
• pp.47-58
• /
• 1987

It has been reported that the luteal function may be regulated by the intracellular $Ca^{++}$ level which may be adjusted partially by the high affinity $Ca^{++}-ATPase$ in luteal cell membranes. Then, one may expect that luteotropic and/or luteolytic agents, such as gonadotropins, prostaglandin $F_{2{\alpha}}\;(PGF_{2{\alpha}})$ and ouabain, affect the intracellular $Ca^{++}$ level. In this present study, therefore, we examined the effects of luteinizing hormone (LH, or human chorionic gonadotropin, hCG), $PGF_{2{\alpha}}$ and ouabain on the kinetic properties of the high affinity $Ca^{++}-ATPase$ in light membrane, heavy membrane, and microsomal fractions from the highly luteinized ovary. LH (or hCG) increased the affinity and the Vmax for $Ca^{++}$ both in light membrane and heavy membrane. $PGF_{2{\alpha}}$ increased the Vmax in light membrane and decreased the Km in heavy membrane for $Ca^{++}$ at low concentration $(5\;{\mu}g/ml)$. At higher concentration, however, $PGF_{2{\alpha}}$ oppositly affected on kinetic properties, that shown at low concentration. Ouabain, a potent inhibitor of $Na^+-K^+-ATPase$, increased the Km at high concentration $(10^{-4}\;M)$, however, decreased the Vmax for $Ca^{++}$ in light membrane at low concentration $(10^{-6}\;M)$. Also, ouabain increased the Km for $Ca^{++}$ in heavy membrane without changes in the Vmax at both concentrations. It seems that LH and low dose of $PGF_{2{\alpha}}$ increase the intracellular $Ca^{++}$ level and cause in activation of $Ca^{++}-ATPase$, however, higher dose of $PGF_{2{\alpha}}$ and ouabain inhibit directly $Ca^{++}-ATPase$ activity and result in increase in intracellular $Ca^{++}$ level. According to the above results, we suggest that luteotropic and/or luteolytic agents regulate the luteal progesterone $(P_4)$ production through two different pathways; one is cyclic adenosine monophosphate (cAMP)-dependent and another is $Ca^{++}-dependent$. Intracellula. $Ca^{++}$ level regulated by the high affinity $Ca^{++}-ATPase$ may affect both pathways in a time-dependent fashion. LH (or hCG) acts on the luteal $P_4$ production via both pathways. The initial step is $Ca^{++}$ dependent, and the late step is cAMP dependent. $PGF_{2{\alpha}}$ and ouabain increase the intracellular $Ca^{++}$ concentration so that basal luteal $P_4$ production is increased and LH-stimulated $P_4$ production is inhibited by the inhibiting LH-dependent adenylate cyclase activity.

• The Korean Journal of Physiology and Pharmacology
• /
• 제2권6호
• /
• pp.725-732
• /
• 1998

In order to elucidate the molecular mechanism of the intracellular $Ca^{2+}$ overload frequently reported from diabetic heart, diabetic rats were induced by the administration of streptozotocin, the membrane vesicles of junctional SR (heavy SR, HSR) were isolated from the ventricular myocytes, and SR $Ca^{2+}$ uptake and SR $Ca^{2+}$ release were measured. The activity of SR $Ca^{2+}-ATPase$ was $562{\pm}14$ nmol/min/mg protein in control heart. The activity was decreased to $413{\pm}30$ nmol/min/mg protein in diabetic heart and it was partially recovered to $485{\pm}18$ nmol/min/mg protein in insulin-treated diabetic heart. A similar pattern was observed in SR $^{45}Ca^{2+}$ uptakes; the specific uptake was the highest in control heart and it was the lowest in diabetic heart. In SR $^{45}Ca^{2+}$ release experiment, the highest release, 45% of SR $^{45}Ca^{2+}$, was observed in control heart. The release of diabetic heart was 20% and it was 30% in insulin-treated diabetic heart. Our results showed that the activities of both SR $Ca^{2+}-ATPase$ and SR $Ca^{2+}$ release channel were decreased in diabetic heart. In order to evaluate how these two factors contribute to SR $Ca^{2+}$ storage, the activity of SR $Ca^{2+}-ATPase$ was measured in the uncoupled leaky vesicles. The uncoupling effect which is able to increase the activity of SR $Ca^{2+}-ATPase$ was observed in control heart; however, no significant increments of SR $Ca^{2+}-ATPase$ activities were measured in both diabetic and insulin-treated diabetic rats. These results represent that the $Ca^{2+}$ storage in SR is significantly depressed and, therefore, $Ca^{2+}-sequestering$ activity of SR may be also depressed in diabetic heart.

• 한국식품과학회지
• /
• 제18권6호
• /
• pp.443-449
• /
• 1986

근원섬유구성단백질의 SDS-polyacrylamide gel 전기 영동상으로 부터 돼지근육의 red muscle과 while muscle의 근원섬유단백질사이에는 30K성분함량의 특징적 차이가 나타났으며, 생물활성에서도 red muscle쪽이 white muscle쪽보다 높은 ATPase 활성을 나타내었다. 근원섬유단핵질의 열안정성은 D값에서 확실한 차이를 보여 white muscle쪽이 red muscle쪽보다 높은 열안정성을 나타냈고, 열역학량에서도 근섬유 type간의 차이를 보였다. 한편 근원섬유단백질의 열안정성은 생체조직에 가까운 형태일수륵 안정하다는 사실도 확인되었다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제6권2호
• /
• pp.101-107
• /
• 2002

In the present study, the postnatal developmental changes in the expressional levels of cardiac sarcoplasmic reticulum (SR) $Ca^{2+}$ regulatory proteins, i.e. $Ca^{2+}-ATPase,$ phospholamban, and $Ca^{2+}$ release channel, were investigated. Both SR $Ca^{2+}-ATPase$ and phospholamban mRNA levels were about 35% of adult levels at birth and gradually increased to adult levels. Protein levels of both SR $Ca^{2+}-ATPase$ and phospholamban, which were measured by quantitative immunoblotting, were closely correlated with the mRNA levels. The initial rates of $Ca^{2+}$ uptake at birth were about 40% of adult rates and also increased gradually during the myocardial development. Consequently, the relative phospholamban/$Ca^{2+}-ATPase$ ratio was 1 in developmental hearts. $Ca^{2+}$ release channel (ryanodine receptor) mRNA was about $50{\sim}60%$ at birth and increased gradually to adult level throughout the postnatal rat heart development. $^3[H]ryanodine$ binding increased gradually during postnatal myocardial development, which was closely correlated with ryanodine mRNA expression levels during the development except the ryanodine mRNA level at birth. These findings indicate that cardiac SR $Ca^{2+}-ATPase,$ phospholamban, and $Ca^{2+}$ release channel are expressed coordinately, which may be necessary for intracellular $Ca^{2+}$ regulation during the rat heart development.

• Preventive Nutrition and Food Science
• /
• 제5권4호
• /
• pp.230-233
• /
• 2000

The present work was designed to determine whether change in fluidity of the mitochondrial membrane affects mitochondrial {TEX}$F_{1}${/TEX}{TEX}$F_{0}${/TEX}ATPase characteristics in NIDDM-prone BHE/Cdb rat. Isolated mitochondria fom BHE/Cdb rat fed a 6% coconut oil or corn oil were functionally tested by an analysis of its respiration and the coupling of this process to ATP synthesis in presence of oligomycin, a specific inhibitor of oxidative phosphorylation (OXPHOS), that binds to the {TEX}$F_{1}${/TEX}{TEX}$F_{0}${/TEX}ATPase. Mitochondria from rats fed coconut oil were more responsive to the inhibitory action of oligomycin with respect to state 3 respiration, respiratory control (RC) ratio and ADP:P (P/O) ratio than were mitochondria from rats fed corn oil. In state 3 respiration, mitochondria from rats fed coconut oil consumed less oxygen than did mitochondria from rats fed corn oil. RC ratio was lower in the mitochondria from rats fed coconut oil than was mitochondria from rats fed corn oil. In P/O ratio, the mitochondria from rats fed coconut oil had a lower P/O ratio than did mitochondria from rats fed corn oil. The data showed that the chang influidity of the mitochondrial membrane by dietary fat affected mitochondrial {TEX}$F_{1}${/TEX}{TEX}$F_{0}${/TEX}ATPase characteristics. The present study on diet differences in {TEX}$F_{1}${/TEX}{TEX}$F_{0}${/TEX}ATPase characteristics provides considerable insight into the role diets play in the control of mitochondrial function, expecially OXPHOS in NIDDM with mitochondrial defects.

• 한국식품영양과학회지
• /
• 제21권6호
• /
• pp.712-718
• /
• 1992

산 강직근과 알카리 강직근의 사후 자장중 물리, 화학적 변화를 비교 검토하였다. 근섬유의 소편화도는 산강직근이 알카리 강직근에 비하여 저장 3일째부터 월등히 높았다. $Mg^{2+}$-ATPase 활성은 알카리 강직근이 저장 전기간 동안 산 강직근 보다 높았으며, 산 강직근은 저장 1일째, 알카리 강직근은 저장 3일째에 가장 높았다. $Ca^{2+}$-ATPase 활성에 있어서 산 강직근과 알카리 강직근 모두 전 저장기간 동안 약간 증가하는 경향이었으나 산 강직근은 알카리 강직근 보다 전기간 동안 활성도가 낮았다. Actomyosin의 용해성이나 myofibril의 전기영동상은 강직 형태에 따른 유의한 차이가 없었다.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
• /
• pp.340-340
• /
• 1994

Since total hepatic ischemia(IS) occurs with transplantation, there has been interest in evaluating hepatic function after ischemia and subsequent reflow of blood. Four groups of animals were studied: group 1 (sham), group 2 (30mins IS), group 3 (60mins IS), and g.cup 4 (90mins IS). Serum transaminase(STA), wet weight-to-dry weight ratio(W/D), lipid peroxides(LPO), glucose-6-phosphatase(G-6-Pase) activity, Na$\^$+//K$\^$+/-ATPase(ATPase) activity were measured at 1, 5 and 24hrs after hepatic ischemia. Significant changes occurred between 1 and 5hrs of reperfusion. STA was 3579${\pm}$401, 4593${\pm}$675 and 6348${\pm}$808 U/L in group 2, 3 and 4 respectively. These changes were ischemic time-dependent manner. W/D in group 3 and 4 were significantly increased than that in sham group at all time points measured. In sham group, the level of LPO in the liver microsome remained constant at approximately 0. 5nmole MDA formed/mg protein througllout the experiment, In all ischemic groups on the other hand, the level of LPO started to increase at ischemia and markedly increased at all reperfusion period. Similar to STA, these changes were also dependent on duration of ischemia. Although G-6-Pase activity remained unchanged in both group 2 and group 3 until 5hrs of reperfusion, marked decrease in G-6-Pase activity was observed at grcup 4. ATPase activity was significantly decreased at 1, 5 and 24 hrs of reperfusion in group 3, whereas it was not changed in group 2. Furthermore, ATPase activity in group 4 started to decrease at ischemia and markedly decreased for entire reperfusion period. These data suggest that severity of hepatocellular injury is associated with period of ischemia as well as period of reperfusion.