• Title/Summary/Keyword: sulfhydryl concentration

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Action of Theobromine on Sodium-Potassium activated ATPase in Red Cell Membrane (Theobromine이 적혈구막의 NaK ATPase의 활성도에 대한 작용)

  • Koh, Il-Sup
    • The Korean Journal of Physiology
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    • v.12 no.1_2
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    • pp.25-34
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    • 1978
  • The action of theobromine on the sodium plus potassium activated ATPase activity In the rabbit red cell membrane has teen investigated and the experiments were also designed to determine the mechanism of action of theobromine on the ATPase activity. The following results were observed. 1. The activity of the NaK ATPase from red fell membrane is stimulated by theobromine, and the concentration of theobromine for maximal activity is about 3mM. 2. The activating effect of theobromine on the ATPase, with a given concentration of potassium in the medium, is increased by raising the sodium concentration but activity ratio is decreased. 3. The activating effect of theobromine on the ATPase, with a given concentration of sodium in the medium. is increased by the raising the potassium concentration but activity ratio is decreased. 4. The NaK ATPase activity is increased by small amounts of calcium but decreased by larger amounts. The activity of the enzyme by theobromine is increased by small amounts of calcium but decreased by larger amounts. 5. The activating effect of theobromine on the ATPase was not related to the hydroxyl group of threonine and imidazole group of histicline. 6. The activating effect of theobromine on the ATPase is due to sulfhydryl group, amino group and carboxyl group of the enzyme of NaK ATPase.

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Action of Pilocarpine on Sodium-Potassium activated ATPase in Rabbit Red Cell Membrane (Pilocarpine이 토끼 적혈구막의 NaK ATPase의 활성도에 대한 작용)

  • Koh, Il-Sup
    • The Korean Journal of Physiology
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    • v.11 no.1
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    • pp.11-20
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    • 1977
  • The action of pilocarpine on the sodium plus potassium activated ATPase activity in the rabbit red cell membrane has been investigated and the experiments were also designed to determine the mechanism of action of pilocarpine on the ATPase activity. The following results were observed. 1. The activity of the NaK ATPase from red cell membrane is stimulated by pilocarpine, and the concentration of pilocarpine for maximal activity is about 3 mM. The pH optimum for the pilocarpine sensitive component is 8.0. 2. The activating effect of pilocarpine on the ATPase, with a given concentration of sodium .in the medium, is increased by raising the potassium concentration but activity ratio is decreased 3. The activating effect of pilocarpine on the ATPase, with a given concentration of Potassium in the medium, is increased by raising the sodium concentration but activity ratio is decreased 4. The NaK ATPase activity is increased by small amounts of calcium but decreased by 'larger amounts. The activity ratio of the enzyme by pilocarpine is decreased by small amounts .of calcium but decreased by larger amounts. 5. The activating effect of pilocarpine on the ATPase was not related to the sulfhydryl group of cysteine, the hydroxyl group of threonine or the imidazole group of histidine. 6. The activating effect of pilocarpine on the ATPase is due to amino group and carboxyl group of the enzyme of NaK ATPase

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Action of Anthraquinone on Sodium-Potassium activated -ATPase in Rabbit Red Cell Membrane- (Anthraquinone이 토끼 적혈주막의 NaK ATPase웨 활성도에 대한 작용)

  • Koh, Il-Sup
    • The Korean Journal of Physiology
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    • v.11 no.1
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    • pp.1-9
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    • 1977
  • Action of anthraquinone on the sodium plus potassium activated ATPase activity in the rabbit red cell membrane has been investigated and the experiments were also designed to determine the mechanism of action of anthraquinone on the ATPase activity. The following results were obtained 1. The activity of the NaK ATPase from red cell membrane is inhibited by anthraquinone and the concentration of anthraquinone for maximal inhibition is about 5mM. 2. The ratio of inhibition of NaK ATPase by anthraquinone, with a giving concentration of sodium in the medium, is increased by raising the potassium concentration. 3. The ratio of inhibition of NaK ATPase by anthraquinone, with a given concentration of potassium in the medium, is increased by raising the sodium concentration. 4. The action of anthraquinone on the NaK ATPase activity is inhibited by calcium ions and the ratio of inhibition is increased by small amounts of calcium but almost constant by larger amounts. 5. The inhibitory action of anthraquinone on the NaK ATPase activity was not related to the amino group of lysine, the hydroxyl group of threonine or the imidazole group of histidine. 6. The inhibitory action of anthraquinone on the ATPase activity is due to sulfhydryl group or the carboxyl group of the enzyme of NaK ATPase.

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Cysteine antagonism of captafol induced toxicities in rats 1. Effects on hematological and serum biochemical values (랫트에 있어서 captafol의 독성에 대한 cysteine의 방어 작용 1. 혈액학 및 혈청 생화학적 성상에 미치는 영향)

  • Kim, Sung-hoon
    • Korean Journal of Veterinary Research
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    • v.35 no.3
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    • pp.437-445
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    • 1995
  • This experiment was carried out to study the preventive effect of cysteine on the toxicities of captafol to the hematological and serum biochemical values. A single dose of captafol(5mg/kg BW, ip) was given to male Sprague-Dawley rats and its toxicities were evalutated by body weitht changes, autopsy findings, absolute organ weight, and hematological and serum biochemical parameters. The single dose of captafol caused significant decreases in body weitht, and absolute liver weight, as-cites, fibrin clot in abdominal cavity, adhesion of liver lobes significant elevation of number of RBC, hemoglobin concentration and serum AST activity, and decreased of serum phospholipid level. Where as cysteine(over 58mg/kg BW) given immediately after captafol appeared to prevent the ascites, fibrin clot in abdominal cavity and liver lobe adhesion. It also prevented the liver and blood, especially RBC toxicites. The results suggest that cysteine and other compounds containing sulfhydryl groups may protect the subjects from captafol-induced toxicity.

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Reaction Mechanism of Purine Nucleoside Phosphorylase and Effects of Reactive Agents for SH Group on the Enzyme in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 얻은 Purine Nucleoside Phosphorylase의 반응기작과 효소에 대한 Sulfhydryl Reagent의 영향)

  • Choi, Hye-Seon
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.222-231
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    • 1994
  • Kinetic analysis was done to elucidate the reaction mechanism of purine nucleoside phosphorylase (PNP) in Saccharomyces cerevisiae. The binary complexes of PNP${\cdot}$phosphate and PNP${\cdot}$ribose 1-phosphate were involved in the reaction mechanism. The initial velocity and product inhibition studies demonstrated were consistent with the predominant mechanism of the reaction being an ordered bi, bi reaction. The phosphate bound to the enzyme first, followed by nucleoside and base were the first product to leave, followed by ribose 1-phosphate. The kinetically suggested mechanism of PNP in S. cerevisiae was in agreement with the results of protection studies against the inactivation of the enzyme by sulfhydryl reagents, p-chloromercuribenzoate (PCMB) and 5,5'-dithiobisnitrobenzoate (DTNB). PNP was protected by ribose 1-phosphate and phosphate, but not by nucleoside or base, supporting the reaction order of ordered bi, bi mechanism. PCMB or DTNB-inactivated PNP was totally reactivated by dithiothreitol (DTT) and the activity was returned to the level of 77% by 2-mercaptoethanol, indicating that inactivation was reversible. The kinetic behavior of the PCMB-inactivated enzyme had been changed with higher $K_m$ value of inosine and lower $V_m$, and was restored by DTT. Inactivation of enzyme by DTNB showed similar pattern of K sub(m) value with that by PCMB, but had not changed the $V_m$ value, significantly. Negative cooperativity was not found with PCMB or DTNB treated PNP at high concentration of phosphate.

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Effect of X-Irradiation on the Levels of some Sulfhydryl Groups, Protein and Cell Volume of Ehrlich Ascites Tumour Cells (X-선(線) 조사(照射)가 Ehrlich 암세포(癌細胞)의 용적(容積), 단백양(蛋白量) 및 수종(數種) Sulfhydryl 기(基)에 미치는 영향(影響)에 관(關)하여)

  • Yu, Choon-Shik;Choo, Young-Eun
    • The Korean Journal of Physiology
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    • v.3 no.2
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    • pp.9-16
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    • 1969
  • It is well known that a number of -SH and -SS containing substances afford a certain measure of protection against radiation effects in many biological systems, and it is conceivable that inherent -SH levels in Ehrlich ascites tumour (ELD)cells may be of decisive improtance with respect to the development of cellular radiation injury. So far, little effort has been directed to elucidate the changes in levels of different -SH and -SS groups in ELD cells when the tumour-bearing whole animal was subjected to the sublethal dose of X-irradiation. The present study was designed to bring some lights in the possible changes of and relationship between various sulfhydryl levels, such as P-SH, NP-SH and NP-SS, as well as the content of protein and cell volume of ELD cells, after subjecting the ELD mice to 1,200 r of X-irradiation. The animals used in this experiment were all mixed bred mice of $20{\sim}25\;gm$ in body weight (approximately 2 months old) irrespective of sex. 12 mice in one experiment were inoculated intraperitoneally with 0.2 ml of ascites tumour cells $(2{\times}10^6\;cells)$, and on the 7th day of the tumour growth, they were X-irradiated with 1,200 r, using the conventional X-ray machine under the following conditions: 200 Kv at 15 mA, 0.5 mm Cu filter, target-skin distance: 50 cm. Radiation dose was measured with the the Philip integrating dosimeter. At 24, 36, 48 and 60 hours after the X-irradiation, the mice were killed by cervical dislocation, and the tumours were taken out. Freshly withdrawn ascites tumours were placed in ice, and immediately the cell concentration was measured with the Coulter Cell Counter (Model B), and the hematocrit of the tumour cells were also determined. Cell volume was thus calculated by the cell concentration and hematocrit value. P-SH content of ELD cells was measured potentiometrically according to the method of Calcutt & Doxey, and NP-SH and NP-SS contents were measured spectrophotometrically by the method described by Ellman. Protein content of ELD cells was determined with the Folin phenol reagent by Lowry et al. Altogether, 48 experimental mice were used, and 12 mice with the only exception of X-irradiation were used as the control. Results obtained indicate that the contents of all the cellular sulfhydryl groups as well as cell volume and protein content of the ELD cells increase significantly as time progresses after the sub-lethal X-ray dose of 1,200 r was given and that all the increase is in a lineal fashion. The regression lines of the relative values, (i. e., taking each control value as 1) of all the values obtained, and the regression lines of cell volume, protein and NP-SH are identical, whereas those of NP-SS and P-SH appear to be widely seperated. However, the difference of those two lines (NP-SS & P-SH) were found to be not significant statistically (p>0.05). Therefore, it can be concluded from the above results that all the values examined increase in a lineal fashion with no statistically significant difference among them. Also, with the radiation dose of 1,200 r, the ELD cell becomes enlarged and swollen progressively up to 60 hours post-irradiation and it becomes more than two times of the original normal size at 60 hours after the irradiation, and up to this stage, it seems apparent that the cell division has been slow due to the X-irradiation applied in this experiment. It is well understandable that the contents of NP-SH, NP-SS, P-SH and protein of the ELD cells increase in parallel with the increase of the cell volume by the X-ray does used, but it also seems interesting to note that all the cellular substances tested show no appreciable difference in the pattern of increase.

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Action of Serotonin on Sodium-Potassium Activated ATPase in Rabbit Red Cell Membrane (토끼 적혈구막의 NaK ATPase의 활성도에 대한 serotonin의 작용)

  • Chung, Soon-Tong;Park, Chul-Bin;Koh, Il-Sup
    • The Korean Journal of Physiology
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    • v.10 no.1
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    • pp.25-34
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    • 1976
  • The action of serotonin on the sodium plus potassium activated ATPase activity in the rabbit red cell membrane has been investigated. The experiments were also designed to determine the mechanism of action of serotonin on the ATPase activity. The following results were obtained. 1) The NaK ATPase activity of rabbit red cell ghosts is stimulated by low concentration of serotonin but inhibited by higher concentration, and the concentration of serotonin for maximal activity is about 2mM. The pH optimum for the serotonin sensitive component is 8.0. 2) The activating effect of serotonin on the ATPase, with a given concentration of sodium in the medium, is increased by raising the potassium concentration but the ratio of activity is decreased. 3) The activating effect of serotonin on the ATPase, with a given concentration of potassium in the medium, is increased by raising the sodium concentration but the ratio of activity is decreased. 4) The ATPase activity is increased by small amounts of calcium but inhibited by larger amounts and the ratio of activity by serotonin is decreased by small amounts of calcium but increased by larger amounts. 5) The action of serotonin on the ATPase activity was not related to the amino group of lysine, the hydroxyl group of threonine, the carboxyl group of aspartic acid, or the imidazole group of histidine. 6) The action of serotonin on the ATPase activity is due to sulfhydryl group of the enzyme of NaK ATPase.

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S-nitrosation Ameliorates Homocysteine-mediated Neurotoxicity in Primary Culture of Bat Cortical Neurons (흰쥐 대뇌피질 신경세포에 미치는 호모시스틴의 신경독성에 대한 S-nitrosation의 역할)

  • Kim, Won-Ki
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.169-175
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    • 1996
  • The reactivity of the sulfhydryl (thiol) group of homocysteine has been associated with an Increased risk of atherosclerosis, thrombosis and stroke. Thiols also react with nitric oxide (NO, an endothelium-derived relaxing factor (EDRF) ), forming S-nitrosothiols that have been reported to have potent vasodilatory and antiplatelet effects and been expected to decrease adverse vascular effects of homocysteine. The present study was aimed to Investigate whether the S-nitrosation of homocysteine modulates the neurotoxic effects of homocysteine. An 18 hour-exposure of cultured rat cortical neurons to homocysteine ( >1 mM) resulted in a significant neuronal cell death. At comparable concentrations ( <10 mM), however, S-nitrosohomocysteine did not induce neuronal cell death. Furthermore, S-nitrosohomocysteirle partially blocked NMDA-mediated neurotoxicity. S-nitrosohomocysteine also decreased NMDA-mediated increases in intracellular calcium concentration. The present data indicate that in brain nitric oxide produced from neuronal and nonneuronal cells can modulate the potential, adverse properties of homocysteine.

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Effect of Samhwasan on Na-K-ATPase Activity in Microsomal Fraction of Rabbit Heart Ventricles (삼화산(三和散)이 심장(心臟) Na-K-ATPase 활성(活性)에 미치는 영향(影響))

  • Shin, Hyeon-Chul;Yoon, Cheol-Ho;Jeong, Ji-Cheon
    • The Journal of Korean Medicine
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    • v.17 no.2 s.32
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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.

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Effect of Ginseng on Sodium-Potassium activated ATPase in Rabbit Red Cell Membrane (인삼이 토끼 적혈구막의 $Na^{+}-K^{+}-ATPase$의 활성도에 미치는 영향)

  • Kang, Byoung-Nam;Koh, Il-Sup
    • The Korean Journal of Physiology
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    • v.8 no.1
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    • pp.55-65
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    • 1974
  • The effect of ginseng on the ATPase activity of rabbit ref cell membrane has been investigated. The experiments were also designed to determine whether the components of ginseng could be attributed to the effect on ATPase activity which dependent upon sodium plus potassium and is sensitive to ouabain. The following results were observed. 1. The activity of the $Na^{+}-K^{+}-ATPase$ from red cell membrane is stimulated by ginseng, and the concentration of ginseng for half-maximal activity is about 15 mg%. The pH optimum for the ginseng sensitive component is 7.6. 2. The portion of the enzyme activity stimulated by ginseng is completely abolished by ouabain. 3. The activating effect of ginseng on the ATPase, with a given concentration of sodium in the medium, is increased by raising the potassium concentration but activity ratio is decreased. 4. The activating effect of ginseng on the ATPase, with a given concentration of potassium in the medium, is increased by raising the sodium concentration but the activity ratio is decreased. 5. The ATPase activity is increased by small amounts of calcium but inhibited by larger amounts and the rate of activity by ginseng is constant. 6. The action of ginseng on the ATPase activity was not related to the sulfhydryl group of cysteine, the amino group of lysine, the imidazole group of histidine, the quanidinium group of arginine, the carboxyl group of aspartic acid, or the hydroxyl group of threonine. 7. The activating effect of ginseng on the ATPase activity may be not due to a saponin which is contained in ginseng.

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