• Title/Summary/Keyword: $H^+-ATPase$

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Different Distribution of the ${alpha}_{2},Na^+,K^+-ATPase lsoform between Rat Atria and Ventricles$

  • Lee, Jeung-Soo;Lee, Shin-Woong;Wallick, Earl-T
    • Archives of Pharmacal Research
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    • v.19 no.5
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    • pp.381-385
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    • 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.

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The Effect of Soil Amended with β-glucan under Drought Stress in Ipomoea batatas L. (𝛽-glucan 토양혼합에 따른 고구마의 가뭄피해 저감 효과 )

  • Jung-Ho Shin;Hyun-Sung Kim;Gwan-Ju Seong;Won Park;Sung-Ju Ahn
    • Ecology and Resilient Infrastructure
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    • v.10 no.3
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    • pp.64-72
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    • 2023
  • Biopolymer is a versatile material used in food processing, medicine, construction, and soil reinforcement. 𝛽-glucan is one of the biopolymers that improves the soil water content and ion adsorption in a drought or toxic metal contaminated land for plant survival. We analyzed drought stress damage reduction in sweet potatoes (Ipomoea batatas L. cv. Sodammi) by measuring the growth and major protein expression and activity under 𝛽-glucan soil amendment. The result showed that sweet potato leaf length and width were not affected by drought stress for 14 days, but sweet potatoes grown in 𝛽-glucan-amended soil showed an effect in preventing wilting caused by drought in phenotypic changes. Under drought stress, sweet potato leaves did not show any changes in electrolyte leakage, but the relative water content was higher in sweet potatoes grown in 𝛽-glucan-amended soil than in normal soil. 𝛽-glucan soil amendment increased the expression of plasma membrane (PM) H+-ATPase, but it decreased the aquaporin PIP2 (plasma membrane intrinsic protein 2) in sweet potatoes under drought stress. Moreover, water maintenance affected the PM H+-ATPase activity, which contributed to tolerance under drought stress. These results indicate that 𝛽-glucan soil amendment improves the soil water content during drought and affects the water supply in sweet potatoes. Consequently, 𝛽-glucan is a potential material for maintaining soil water contents, and analysis of the major PM proteins is one of the indicators for evaluating the biopolymer effect on plant survival under drought stress.

Comparative Biochemical Study on the Myofibrillar Proteins from Porcine Muscle (Porcine Myofibrillar Protein에 대한 비교생화학적 연구)

  • Yang, Ryung;Park, Hyun-Joo;Kim, Young-Ho;Jhin, Hong-Seung;Shin, Wan-Chul
    • Korean Journal of Food Science and Technology
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    • v.18 no.6
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    • pp.443-449
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    • 1986
  • In order to investigate the general characteristics of ATPase and ATPase thermostability between porcine white muscle and red muscle, myofibrillar proteins were prepared and compared their physicochemical characteristics. SDS-polyacrylamide gel electrophoretic analyses showed that a protein band of 30,000 daltons was detected noticeably in myofibril from red muscle, but negligibly in myofibril from white muscle. The noticeable differences were found between porcine white muscle and red muscle for the activities of EDTA-ATPase, Ca-ATPase and Mg-ATPase. Myofibrillar proteins from white muscle showed higher thermostability than those from red muscle. Thermodynamic parameters, enthalpy $({\Delta}H^#)$, entropy $({\Delta}S^#)$, etc., showed characteristic variations between porcine white muscle and red muscle.

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Changes in the Expressional Levels of Sarcoplasmic Reticulum $Ca^{2+}-regulatory$ Proteins in the Postnatal Developing Rat Heart

  • Lee, Eun-Hee;Park, Soo-Sung;Lee, Jae-Sung;Seo, Young-Ju;Kim, Young-Hoon;Kim, Hae-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.2
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    • pp.101-107
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    • 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.

Human ChlR1 Stimulates Endonuclease Activity of hFen1 Independently of ATPase Activity

  • Kim, Do-Hyung;Kim, Jeong-Hoon;Park, Byoung Chul;Lee, Do Hee;Cho, Sayeon;Park, Sung Goo
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.3005-3008
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    • 2014
  • Human ChlR1 protein (hChlR1), a member of the cohesion establishment factor family, plays an important role in the segregation of sister chromatids for maintenance of genome integrity. We previously reported that hChlR1 interacts with hFen1 and stimulates its nuclease activity on the flap-structured DNA substrate covered with RPA. To elucidate the relationship between hChlR1 and Okazaki fragment processing, the effect of hChlR1 on in vitro nuclease activities of hFen1 and hDna2 was examined. Independent of ATPase activity, hChlR1 stimulated endonuclease activity of hFen1 but not that of hDna2. Our findings suggest that the acceleration of Okazaki fragment processing near cohesions may aid in reducing the size of the replication machinery, thereby facilitating its entry through the cohesin ring.

Transport of Tetraethylammonium in Renal Cortical Endosomes of Cadmium-Intoxicated Rats

  • Park, Hee-Seok;Kim, Kyoung-Ryong;Park, Yang-Saeng
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.1
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    • pp.21-26
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    • 2002
  • Effects of cadmium (Cd) intoxication on renal endosomal accumulation of organic cations $(OC^+)$ were studied in rats using $^{14}C-tetraethylammnium$ (TEA) as a substrate. Cd intoxication was induced by s.c. injections of 2 mg Cd/kg/day for $2{\sim}3$ weeks. Renal cortical endosomes were isolated and the endosomal acidification (acridine orange fluorescence change) and TEA uptake (Millipore filtration technique) were assessed. The TEA uptake was an uphill transport mediated by $H^+/OC^+$ antiporter driven by the pH gradient established by $H^+-ATPase.$ In endosomes of Cd-intoxicated rats, the ATP-dependent TEA uptake was markedly attenuated due to inhibition of endosomal acidification as well as $H^+/TEA$ antiport. In kinetic analysis of $H^+/TEA$ antiport, Vmax was reduced and Km was increased in the Cd group. Inhibition of $H^+/TEA$ antiport was also observed in normal endosomes directly exposed to free Cd (but not Cd-metallothionein complex, CdMt) in vitro. These data suggest that during chronic Cd exposure, free Cd ions liberated by lysosomal degradation of CdMt in proximal tubule cells may impair the endosomal accumulation of $OC^+$ by directly inhibiting the $H^+/OC^+$ antiporter activity and indirectly by reducing the intravesicular acidification, the driving force for $H^+/OC^+$ exchange.

SOME PHYSICO-CHEMICAL CHARACTERISTICS OF CARP MYOSIN B (잉어 Myosin B의 몇 가지 생물물리화학적인 성질)

  • PYEUN Jae Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.5 no.2
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    • pp.57-62
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    • 1972
  • Natural actomyosin was extracted from the dorsal muscle of carp (Cyprinus carpio), and some Physico-chemical properties were determined. The sedimentation constant ($s_{20},\;_\omega$) of the purified skeletal muscle actomyosin was about 24.76s and the ATP sensitivity 147-176. The specific viscosity was rapidly ascended with the increase of protein concentration as other vertebrate actomyosins. ATPase activating site under the effect of pH variation was showed on the both sides of acid and base resembling with the tendency of rabbit skeletal muscle actomyosin, and found at 6.0 on the acidic side and at 9.5 on the basic side.

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Characterizing Salt Stress Response in a Rice Variety and Its Salt Tolerant Lines Derived from In Vitro Mutagenesis

  • Lee In Sok;Kim Dong Sub;Kang Si Yong;Wi Seung Gon;Jin Hua;Yun PiI-Yong;Lim Yong Pyo;Lee Young Il
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.205-212
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    • 2004
  • The objectives were to compare the salt tolerance levels in the parental rice cultivar, Dongjinbyeo, and induced mutagenesis derived its lines for plant height, MDA, ATPase, POD, and 2-dimensional protein electrophoresis pattern in NaCl-containing hydroponic nutrient solutions. Rice plants isolated from a population of rice (Oryza sativa L. cv. Dongjinbyeo) mutation lines, which were generated in combination with in vitro selection and gamma-ray, exhibited salt tolerance. Line No. 18 had the longest plant, whereas NaCl-sensitive line (No. 25) had the shortest plant. The parent, and the sensitive line showed severe damage from salt stress. Tolerant lines (No. 18, 50) had a lower malonaldehyde (MDA) content than the sensitive one (Dongjinbyeo, No. 25) during salt stress. Several proteins showed significant quantitative variation through 2DE; phosphoribulokinase, peroxidase, oxygen evolving enhancer 1 and the $H^+-ATPase$, which are known to be involved in salt tolerance. The effect of salt on peroxidase and $H^+-ATPase$ activity in the seedlings of two groups with contrasting genotypes of rice was studied. A greater activity was recorded in the tolerant lines as compared to the sensitive ones (P<0.05, Duncan's test). The results indicate that salt tolerant lines expressed more salt stress-inducible proteins associated with salt tolerance than the sensitive lines during salt stress.

Study on the Role of Metal ions for the Activity of the Mitochondrial $F_1-ATPase$ in Lentinus edodes (표고버섯의 Mitochondrial $F_1-ATPase$ 활성도에 미치는 금속이온의 역할에 관한 연구)

  • Park, Sang-Shin;Min, Tae-Jin
    • The Korean Journal of Mycology
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    • v.22 no.2
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    • pp.122-129
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    • 1994
  • The role of metal ions for the activity of the mitochondrial $F_1-ATPase$ was studied. Removal of non-heme iron ion from the mitochondria by dialysis against chelating agents, 10 mM ethylenediaminetetraacetic acid(EDTA) and 10 mM o-phenanthroline(o-Phe), led to 56% and 49% inactivation of the enzyme, respectively. The enzyme dialyzed against EDTA was reactivated 81% by the addition of 0.5 mM $Fe^{3+}$ and 70% by 0.5 mM $Mg^{2+}$. But, $Fe^{2+}$ did not reactivate the enzyme. Coexistence of 0.5 mM $Fe^{2+}$ and 0.5 mM $Mg^{2+}$ resulted in 95% reactivation of the enzyme, while $Fe^{3+}$ with 0.5 mM $Mg^{2+}$ did not reactivate the enzyme like the effect of $Fe^{2+}$ alone. The enzyme dialyzed against o-Phe showed the similar results. These data showed that $Fe^{3+}$ is predominantly required for the activity of the mitochondrial $F_1-ATPase$ in Lentinus edodes and stimulated the activity of it by $Mg^{2+}$. $Fe^{3+}$ and $Mg^{2+}$ increased enzyme's affinity for substrate, decreasing the Km value 1.67 mM to 0.65 mM.

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Aluminum-induced Root Growth Inhibition and Impaired Plasma Membrane $H^+-flux$ in Mung Bean (알루미늄에 의한 녹두 뿌리의 생장 억제와 원형질막 $H^+-flux$의 손상)

  • Ahn, Sung-Ju;Kim, Yu-Sun;Park, Won;Ku, Yang-Gyu;Min, Kyung-Soo;Whang, Tei-Ik
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.2
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    • pp.213-219
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    • 2007
  • It has been well established that aluminum (Al) inhibits root tip growth rapidly in acid soil. We report the correlation between Al induced growth inhibition and impaired $H^+-flux$ in mung bean (Vigna radiate L. cv. Kumsung). The root growth inhibition was dependent on Al concentration (0, 10, 25, 50, $100{\mu}M$) and exposure time (12 and 24 h). Using Hematoxylin staining, it was observed that the root damage was occurred preferentially in regions with high Al accumulation. Using the pH indicator, it was shown that the surface pH of root tip was strongly alkalized in the control whereas changed only slightly in the $50{\mu}M$ Al-treated root. The $H^+-ATPase$ activity of plasma membrane vesicles was inhibited by 56% in the Al-treated roots compared to control root. Decrease in the amount of the plasma membrane $H^+-ATPase$ (100 kDa) translation in the plant roots under Al stress was demonstrated by Western blot analysis. These results indicate that the dynamics of $H^+-flux$ across the root tip play an important role in root growth under Al stress.