• The Korean Journal of Mycology
• /
• v.17 no.4
• /
• pp.169-176
• /
• 1989

Mitochondria in Pleurotus ostreatus was purified by stepped sucrose density gradient centrifugation. The activity of mitochondrial ATPase has been investigated during various times of illumination at each wavelength in the range of 400 nm to 700 nm. The mitochondrial ATPase activity was simulated 1,7 fold by 580 nm illumination compared with the broad wavelength group. The mitochondrial ATPase activity according to various times of illumination was stimulated 2.2 fold for 10 seconds at 580 nm compared with the broad wavelength group. The optimum pH and temperature of the mitochondrial ATPase were 7.4 and $60^{\circ}C$, respectively. The activity of this enzyme was stimulated by 5 mmol $Fe^{3+}$, 5 mmol $Mg^{2+}$, 0.1 mmol $Ca^{2+}$ and 5 mmol $Fe^{2+}$ ion, but inhibited by 5 mmol $Na^{+}$ ion.

• 한국유가공학회:학술대회논문집
• /
• 1997.05a
• /
• pp.23-34
• /
• 1997

Calcium-stimulated ATPase ($Ca^{2+}$-ATPase) which has optimal pH value at 7.0 was found in the membrane fraction of human milk, and its enzymatic properties were studied. The purified $Ca^{2+}$-ATPase required 0.45 mM Ca ion for maximal activity. Among the nucleosides, $Ca^{2+}$-ATPase showed a higher substrate specificity to ATP and UTP than to CTP and GTP. $Ca^{2+}$-ATPase had apparent Km value of 0.065, and V max of 7.63 mol ATP hydrolyzed/mg pro-tein per min, respectively. $Ca^{2+}$-ATPase was potently inhibited by lanthanide, vanadate, and p-chloromercuribenzoate, and inactivated by EDTA, and CDTA and EGTA, but were unaffected by N-ethylmaleimide, $NaN_3$, ouabain, or oligomycin, and was completely inactivated by heating at $60^{\circ}C$ for 10 min. This enzyme activity was concentrated in the membrane fraction of the cream and skim milk membrane, but not founded in bovine milk.

• Environmental Mutagens and Carcinogens
• /
• v.8 no.1
• /
• pp.35-46
• /
• 1988

출생직후부터 새끼쥐에 유발시킨 납중독이 중추의 특정 신경계에 미치는 신경독성의 선택성 여부를 알아보고자 하였다. 특정 신경계의 한 예로 모노아민성 신경계를 선택하여 납중독의 지표로 모노아민성 신경계의 효소인 MAO (mono-amine oxidase)의활성을 측정하였으며, 비특정 조직의 지표로 Na+K+-ATPase의 활성을 측정하였다. Wistar계의 흰쥐에서 태어난 새끼들에게 출생직후부터 전실험기간을 통해 0.05% 혹은 0.2% 초산납 (PbAc2)용액을 식수로 공급하여 납중독을 유발시켰다. 생후 2, 4, 6 및 8주된 새끼쥐의 MAO 및 Na+K+-ATPase활성을 대뇌, 간뇌, 중뇌, 뇌교-연수 및 다섯부위에서 각기 측정하였다.

• Environmental Mutagens and Carcinogens
• /
• v.7 no.2
• /
• pp.72-84
• /
• 1987

어미쥐에 유발시킨 납중독이 새끼쥐의 특정 중추신경계에 미치는 신경독성의 선택성 여부를 알아보고자 하였다. 특정 신경계의 한 예로 모노아민성 신경계를 선택하여 납중독의 지표로 모노아민성 신경계의 효소인 MAO(monoamine oxidase)의 활성을 측정하였으며 비특정조직에의 지표로 Na+.K+-ATPase의 활성을 측정하였다. 임신한 Wistar계 어미쥐에게 임신전기간에 걸쳐 0.05 혹은 0.2% 초산납(PbAc2)용액을 식수로 공급하여 간접적으로 태아에 납중독을 유발시켰다. 새끼쥐는 출생직후 정상 식수를 공급해 주었다. 2, 4, 6 및 8주된 새끼쥐의 MAO 및 Na+.K+-ATPase활성을 대뇌, 간뇌, 중뇌, 뇌교-연수 및 소뇌 등 다섯부위에서 각기 측정하였다.

• Applied Biological Chemistry
• /
• v.41 no.2
• /
• pp.130-136
• /
• 1998

Microsomes of tomato roots were prepared and the activities of microsomal ATPases were measured in order to understand the molecular mechanisms of various ion transports. The activities of plasma membrane $H^+-ATPase$ and vacuolar $H^+-ATPase$ were evaluated to ${\sim}30%$ and ${\sim}38%$ of total microsomal ATPase activity by using their specific inhibitor, vanadate and nitrate $(NO^-_3)$, respectively. The inhibitory effects of vanadate and $NO^-_3$ were additive and the simultaneous additions of these two inhibitors decreased the total activity up to $50{\sim}70%$. The microsomal ATPase activity was regulated key pH and the maximal activity was obtained at pH 7.4. The activity of microsomal ATPase was increased by $K^+$ up to ${\sim}30%$ at the concentration of $K^+$ above 10 mM. However, the $K^+-induced$ increase in the activity was completely inhibited by the simultaneous addition of $Na^+$. To identify the ATPase activity regulated by $K^+$, the effects of specific inhibitors were measured. Vanadate and $NO^-_3$ inhibited total ATPase activity by 27% and 32% in the absence, of $K^+$ and by 27% and 40% in the presence of 120 mM $K^+$, respectively. These results suggest that $K^+$ increases the activity of $NO^-_3-sensitive$ vacuolar $H^+-ATPase$ but not that of vanadate-sensitive plasma membrane $H^+-ATPase$ since vanadate has no effect on $K^+-induced$ increase in ATPase activity. The microsomal ATPase activity was also decreased by increasing $Ca^{2+}$ concentration. Interestingly, $NO^-_3$ blocked the $Ca^{2+}-induced$ inhibition of microsomal ATPase activity; however, vanadate had no effect. These results imply that vacuolar $H^+-ATPase$ is activated by $K^+$ and inhibited by $Ca^{2+}$.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.17 no.4
• /
• pp.271-279
• /
• 1984

Effects of temperature and additives on the stability of actomyosin extracted from skeletal muscle of Israeli carp, Cyprinus carpio nudus, were studied by analyzing free SH-group, ATP-sensitivity and Ca-ATPase activity. The used additives were sucrose, sorbitol, Na-glutamate and L-cysteine. Furthermore, the denaturation constant($K_D$), protective effect(${\Delta}E/M$) and the other thermo-dynamic parameters on protein denaturation are systematically discussed. The actomyosin showed $4.12{\sim}4.68 mg/ml$ in protein concentration, $2.63{\sim}2.93\%$ in ribonucleic acid to the protein, $1:2.20{\sim}2.63$ in the binding ratio of myosin and actin, $4.33{\sim}5.26\%$ in fat content, 109.78 in ATP-sonsitivity, $0.159{\sim}0.201\;{\mu}M-Pi/min/mg-protein$ in Ca-ATPase activity and $3.3{\sim}3.4M/10^5$g-protein in free SH-group content. The first-order rate plots were obtained on the decrease of Ca-ATPase activity and ATP-sensitivity with an increase in temperature, while the free SH-group was increased to $60^{\circ}C$ and decreased rapidly above the temperature. The half-life of Ca-ATPase activity on the actomyosin Ca-ATPase was 280 min at $12^{\circ}C$, 125 min at $20^{\circ}C$, 55 min at $30^{\circ}C$ and 13 min at $40^{\circ}C$, and activation energy, activation enthalpy, activation entropy and free energy of the proteins at $20^{\circ}C$ wene 5,395 cal/mole, 4,814 cal/mole, -40.42 e.u. and 17,626 cal/mole, respectively. The protective effect of the additives on the actomyosin Ca-ATPase showed that the most effective material is $3\%$ sorbitol and followed in the order of $8\%$ Na-glutamate, $1\%$ sucrose and $1\%$ L-cysteine. The actomyosin was more stable at $-30^{\circ}C$ than at $0^{\circ}C$ and $-20^{\circ}C$. and when the additives were used in the low temperature storage, $8\%$ Na-glutamate was the most effective. $3\%$ sorbitol, $1\%$ sucrose and $1\%$ L-cysteine was to become lower in the order.

• Archives of Pharmacal Research
• /
• v.19 no.5
• /
• pp.381-385
• /
• 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 and Pharmacology
• /
• v.12 no.6
• /
• pp.331-336
• /
• 2008

The present study was aimed to determine whether there is an altered regulation of tubular transporters in gentamicin-induced nephropathy. Sprague-Dawley male rats ($200{\sim}250\;g$) were subcutaneously injected with gentamicin (100 mg/kg per day) for 7 days, and the expression of tubular transporters was determined by immunoblotting and immunohistochemistry. The mRNA and protein expression of OAT was also determined. Gentamicin-treated rats exhibited significantly decreased creatinine clearance along with increased plasma creatinine levels. Accordingly, the fractional excretion of sodium increased. Urine volume was increased, while urine osmolality and free water reabsorption were decreased. Immunoblotting and immunohistochemistry revealed decreased expression of $Na^+/K^+$-ATPase, NHE3, NBC1, and AQP1 in the kidney of gentamicin-treated rats. The expression of OAT1 and OAT3 was also decreased. Gentamicin-induced nephropathy may at least in part be causally related with a decreased expression of $Na^+/K^+$-ATPase, NHE3, NBC1, AQP1 and OAT.

• The Korean Journal of Physiology and Pharmacology
• /
• v.10 no.2
• /
• pp.65-69
• /
• 2006

The present study was undertaken to explore the role of autonomic nerves in the regulation of sodium transporters and water channels in the salivary gland. Rats were denervated of their sympathetic or parasympathetic nerves to the submandibular gland. One week later, the expression of Na,K-ATPase, epithelial sodium channels (ENaC), and aquaporins (AQP) was examined in the denervated and contralateral glands. The sympathetic denervation slightly but significantly decreased the expression of ${\alpha}1$ subunit of Na,K-ATPase, whereas the parasympathetic denervation increased it. The expression of ${\alpha}$-subunit of ENaC was significantly increased in both the denervated and contralateral glands either by the sympathetic or parasympathetic denervation. The sympathetic denervation significantly increased the expression of AQP5 in both the denervated and contralateral glands, whereas the parasympathetic denervation decreased it. It is suggested that the autonomic nerves have a tonic effect on the regulation of sodium transporters and AQP water channels in the salivary gland.

• YAKHAK HOEJI
• /
• v.41 no.5
• /
• pp.547-553
• /
• 1997

The purpose of this investigation was to determine the inhibition on $Na^+,\;K^+$-ATPase, cyclic AMP phosphodiesterase and platelet activation by marine sterols isolated from octocorals. Three marine polyhydroxysterols, 7${\alpha},\;8{\alpha}-epoxy-3b{\beta},\;5{\alpha},\;6{\alpha}-trihydroxycholestane (1),\;24-methyl-7{\alpha},\;8{\alpha}-epoxy-3{\beta},\;5{\alpha},\;6{\alpha}-trihydroxycholest-22-ene (2),\;and\;7{\alpha},\;8{\alpha}-epoxy-3{\beta},\;5{\alpha},\;6{\alpha}-trihydroxycholest-22-ene (3)$, were isolated from the Gorgonian Acabaria undulata. Five marine sterols(compound 4, 5, 6, 7, 8) were isolated from the soft coral Alcyonium gracillimum. Three marine polyhydroxysterols (1, 2, 3) and pregna-1. 20-diene-3-one (8) exhibit a potent inhibitory effect on rabbit platelet aggregation induced by collagen and thrombin. Those polyhydroxysterols also exhibit a potent inhibitory effect on cyclic AMP phosphodiesterase. Compound 6 with an unusual cyclic enolether exhibit a inhibitory effect on $Na^+,\;K^+$-ATPase.