골격근 근형질세망의 ATPase활성도에 대한 Vanillylnonanamide의 영향

박경섭,홍장희,류영수,성지연,허강민,임종호,이재흔,석정호 충남대학교 의과대학 지역사회의학연구소 1999 충남의대잡지 Vol.26 No.2

To investigate the effect of vanillylnonanamide(VN) on the ATPase activity of the sarcoplasmic reticulum(SR) of the skeletal muscle, we prepared the SR vesicles from the back muscle of the rabbit, and measured ATPase activity. The results as follows: Ca-ATPase activity was about 50% in the total ATPase activity of skeletal muscular SR. In the reaction mixture with calcium, 100μM VN increased ATPase activity to 20%, and 100 nM and 10 μM thapsigargin(THP) inhibited the ATPase activity to 50% and 60%, respectively. And 100 μM VN plus 100 nM or 10 μM THP more inhibited ATPase activity than THP alone did it. However, without calcium, 100μM VN did not affect ATPase activity, and 10 μM THP inhibited it to 41%, but VN plus THP inhibited it to 31%. The above results show that VN in the reaction mixture with or without calcium has the different action to ATPase activity when it is used alone or together with Ca-ATPase inhibitor THP. This suggests that VN might increase or decrease the skeletal SR Ca-ATPase activity through affecting the lipid membrane around the Ca-ATPase.

Thallium-201 을 이용한 세포막 Na+-K+ ATPase 활성도 측정 ; Rubidium-86 측정법과의 비교

이규보(Kyu Bo Lee),이재태(Jae Tae Lee),손상균(Sang Kyun Sohn),이인규(In Kyu Lee) 대한핵의학회 1998 핵의학 분자영상 Vol.32 No.2

N/A Purpose: Na+-K+ ATPase Activity has beem estimated by the degree of inhibition of cation transport by cardiac glycosides (ouabain) using Rb-86 as a substrate. The biological characterist- Isc of T1-201 is known to be simiIar to those of potassium as a transport substrate in the presence of glucose, insulin or phobol myristate acetate (PMA). The purpose of this study was to measure ouabain sensitive Na+-K+ ATPase activity using T1-201 and compare with that using Rb-86. Materials and Methods: Smooth muscle cells isolated from rat aorta or human placental umbilical artery were cultured, and used to measure cellular Na+-K+ ATPase activity. Na+-K+ ATPase activity was measured as a percentage decrease in cellular uptake of T1-201 or Rb-86 by ouabain under the presence of glucose, insulin or PMA in media. Results: Na+-K+ ATPase ase activity measured with T1-201, as a transport substrate, was not different from those measured with Rb-86 in rat or human smooth muscle cell preparation. Incubation with high concentration glucose resulted in about 30% decrease in enzyme activity. In contrast, insulin or PMA resulted in 50-70% or 28% increases from baseline activity, respectively. Conclusion: These results suggests that T1-201 could replace Rb-86 in measurement of ouabain sensititive Na+-K+ ATPase activity in vitro. High level of glucose concentration decreased cellular Na+-K+ ATPase activity, but insulin or PMA increased it.

저장기간에 따라 추출된 쇠고기 Actomyosin의 생물활성 변화

정인철,김미숙,강세주 한국식품영양학회 1997 韓國食品營養學會誌 Vol.10 No.3

소의 도체로부터 사태, 갈비 및 등심을 분리하고 8℃에 저장하면서 actomyosin을 추출하여 부위별 저장기간에 따라 추출성 및 ATPase활성을 비교하였다. Actin과 myosin이 유리되어 형성된 actomyosin의 추출성은 저장초기 사태, 갈비 및 등심이 각각 36.74, 72.55 및 56.77㎎/g이었으며, 저장기간에 의한 추출양상은 갈비와 등심이 비슷하였고, 사태는 이들과 다르게 진행되었다. 사태의 Mg- 및 Ca-ATPase활성은 저장 3일까지 상승하다가 6일째 감소하였고, 갈비는 저장기간 동안 비슷하였으며, 등심은 저장기간에 따라 조금씩 낮아지는 경향이었다. 그리고 Mg- 및 Ca-ATPase활성은 사태, 등심 및 갈비의 순으로 크게 나타났다. 사태와 갈비의 EDTA-ATPase활성은 저장기간과 이온강도에 따라 차이를 보였지만 등심은 이온강도가 커짐에 따라 계속 상승하였다. This study was carried out to compare the extractability and ATPase activity of actomyosin extracted shank, rib and loin muscle of beef meat stored at 8℃. The extractability of actomyosin in shank, rib and loin muscle were 36.74, 72.55 and 56.77㎎/g early in the storage, respectively. The extractability of the rib and loin muscle were similar, the shank muscle was processed differently with their. The Mg- and Ca-ATPase activity of the shank muscle rised to 3 days, but decreased the 6th day. And Mg-and Ca-ATPase activity of the rib muscle was similar during storage period, the loin muscle made a slow descent. The strength of Mg- and Ca-ATPase activity showed in the order shank, rib and loin muscle. The EDTA-ATPase activity of the shank and rib muscle was difference according to storage period and ionic strength, but the loin muscle was increase in succession with magnitude of ionic strength.

Valium이 가토뇌피질 Homogenate내 Na-K-ATPase활성에 미치는 동력학적 분석

이상봉 인제대학교 1980 仁濟醫學 Vol.1 No.1

Valium이 가토 대뇌피질 Homogenate 내 Na-K-ATPase에 미치는 영향에 대해 효소 역학적 분석을 통하여 그 작용기전을 관찰한 결과 Valium은 Na+이나 K+의 결합부위에 직접 작용하여 Na-K-ATPase의 활성을 억제할 것으로 사료된다. The Kinetics of inhibition of Na-K-ATPase activity by valium was investigated with homogenate of rabbit cerebral cortex. The results were summarized as the fellows: 1.Inhibition of Na-K-ATPase activity was reversible and showed a dose-dependent manner with an estimated I50 of 0.45 mM. 2.Enzyme activity was protected from inhibition by increasing concentrations of enzyme protein and ATP. 3.Altered pH and activity curves for Na-K-ATPase demonstrated comparable inhibition by valium in buffered acidic, neural and alkaline pH ranges. 4.Kinetic studies of cationic-substrate activation of Na-K-ATPase showed noncompetitive inhibition with respect to substrate and competitive inhibition with respect to Na+ and K+. These results suggest that valium appeared to exert its effects on Na-K-ATPase activity by interacting at Na+ and K+ sites.

연체류의 근원섬유단백질에 관한 연구

신완철,송재철,김영호 한국식품영양학회 1997 韓國食品營養學會誌 Vol.10 No.2

근원섬유단백질의 이온강도에 따른 Ca-ATPase 활성, Mg-ATPase 활성 및 EDTA-ATase 활성은 오징어와 대합에서 그 차이점이 뚜렷하였으며, activity-pH curve에서 오징어 actomyosin의 Ca-ATPase 활성은 biphasic response가 소실되었고 대합의 actomyosin은 미약한 biphasic response가 나타났다. 또한 저농도의 dioxane에 의하여 오징어의 근원섬유단백질은 급격한 활성의 감소를 보였으나 대합의 근원섬유단백질은 활성이 증가되었다. 그리고 에탄올과 메탄올은 오징어와 대합의 myosin 및 HMM에 대하여 저농도에서 활성을 증가시켰다. 한편 NEM으로 근원섬유단백질을 modification시키면 10 exp (-6)M 이하의 NEM 농도에서는 활성이 증가되었으나 10 exp (-5)M 이상의 농도가 되면 활성의 급격한 감소가 나타났다. In order to compare and examine the general characteritics of myofibrillar proteins which is an important protein source as a food resource and relates directly with muscle contraction, we have extracted the myofibrillar proteins from squid and clam. The ionic strength of myofibrillar proteins connected with Ca-ATPase activity Mg-ATPase activity and EDTA-ATPase activity showed distinct differences between squid and clam. In the activity-pH curve, actomyosin of the clam had a weak biphasic response. In the low concentration of dioxane, myofibrillar proteins of the squid showed a sudden decrease in activity but myofibrillar proteins of the clam showed in increase in activity. Ethanol and methanol in low concentration caused myosin and HMM from the squid and clam to increase their activities. If we cause modification by NEM, under 10 exp (-6)M concentration, the activity was increased but above 10 exp (-5)M concentration, there was a sudden decrease in activity.

Gonadotropins, Prostaglandin $F_{2{\alpha}}$ 및 Ouabain이 황체막의 $Ca^{++}-ATPase$ 활성도에 미치는 영향

구본숙,김인교,Koo, Bon-Sook,Kim, In-Kyo 대한생리학회 1987 대한생리학회지 Vol.21 No.1

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.

Effect of Cathepsin D Treatment on ATPase Activity of Rabbit Myofibril

YANG, Ryung 朝鮮奬學會 1973 學術論文集 Vol.3 No.-

The effect of cathepsin D and pepsin treatment on rabbit myofibril was studied by measur¬ing the amount of proteolytic products and Mg-enhanced ATPase activity. When myofibril was treated with cathepsin D at 3℃ and pH 5.0 or 5.5, a little but detectable amount of nonprotein nitrogenous compounds was released. However, there was no change in ATPase activity of myofibril, though treated with cathepsin D of higher units than assumed to be in muscle. When myofibril was treated with pepsin under the same condition as used above, there was an increase in KCI-concentration dependence of ATPase activity followed by a decrease in the maximal value of ATPase activity. From the present results, it was concluded that cathepsin D might not take a main role on the post-mortem degradation ofmyofibtil.

Gonadotropins, Prostaglandin F_2α 및 Ouabain이 황체막의 Ca⁺⁺}-ATPase 활성도에 미치는 영향

구본숙(Koo, Bon-Sook),김인교(Kim, In-Kyo) 대한생리학회 1987 대한생리학회지 Vol.21 No.1

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_2α (PGF_2α) 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^2α 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^2α increased the Vmax in light membrane and decreased the Km in heavy membrane for Ca⁺⁺ at low concentration (5 μg/ml). At higher concentration, however, PGF^2α 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^2α increase the intracellular Ca⁺⁺ level and cause in activation of Ca⁺⁺-ATPase, however, higher dose of PGF^2α 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 (P₄) 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 P₄ production via both pathways. The initial step is Ca⁺⁺ dependent, and the late step is cAMP dependent. PGF^2α and ouabain increase the intracellular Ca⁺⁺ concentration so that basal luteal P₄ production is increased and LH-stimulated P₄ production is inhibited by the inhibiting LH-dependent adenylate cyclase activity.

Effect of Verapamil on the Specific Activity of Na-K-activated Adenosine Triphosphatase in Rabit Renal Medulla

Park, Jung Ran,Baik, Haing Woon,Baik, Hyung Hwan,Cho, Yong Ho 慶熙大學校 醫科大學 1990 慶熙 醫大 論文集 Vol.15 No.1

Thermoplasma acidophilum의 20S Proteasome에 의한 ATPase-활성적 단백질분해에 관한 연구

우기민,장예진,조만희,김창세,김완종,조성호,이상한 순천향의학연구소 1999 Journal of Soonchunhyang Medical Science Vol.5 No.1

The eukaryotic 26S proteasome is an ATP/ubiquitin-dependent proteolytic complex consisting of the 20S core particle and 19S ATPase complex. However, because of its complexity and unstable properties, this study was carried out to present more simple and stable model for the ATP-activated proteolytic complex in prokaryotes which can take the place of the eukaryotic 26S proteasome. For this purpose, recombinant Thermoplasma 20S proteasome (T20S) and Methanococcus MS4, a sequence homolog of one ATPase subunit in the 19S ATPase complex, were successfully isolated from Escherichia coli (E. coli). The α and β subunits of T20S expressed in E. coli could assemble for themselves, and showed the peptide-hydrolyzing activity. Whereas both T20S and R20S (the 20S complex from rabbit skeletal muscle) had the highest peptidase activity against Suc-LLVY-AMC, a good substrate for chymotrypsin-like peptidase activity, the specific activity of T20S was slightly lower than that of R20S. In addition, several reagents such as KCI, SDS, and ovalbumin were shown to have different effects on the peptidase activities between T20S and R20S. When the ATPase activity of the purified MS4 were assayed , the Km for ATP was about 0.5 mM, and casein could stimulate the activity more than 2 fold without the change in Km. This result implicates that protein-activated ATPase may induce the conformational change of casein, and therefore suggests that MS4 ATPase may activate the proteolytic activity of the 20S proteasome via accelerating the recognition and translocation of the protein substrates.