• 제목/요약/키워드: $^{45}Ca^{2+}$ uptake

검색결과 69건 처리시간 0.031초

Enzymatic Properties of Low Affinity Vanadate-sensitive ATPase in the Microsomes of Tracheal Epithelial Cells

  • Jung Sakong;Kim, Young-Kee
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1998년도 학술발표회
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    • pp.29-29
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    • 1998
  • Previously, we reported two types of vanadate-sensitive ATPases in the micro somes of tracheal epithelial cells, a high-affinity one and a low-affinity one. The low affinity vanadate-sensitive (LAVS) ATPase was sensitive to thapsigargin and cyclopiazonic acid, specific antagonists of ER-type Ca$\^$2+/-ATPase, and mediated microsomal $\^$45/Ca$\^$2+/ uptake, implying that the LAVS-ATPase is an ER/SR-type Ca$\^$2+/-ATPase.(omitted)

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Characterization of the Ryanodine Receptor and SERCA in Fetal, Neonatal, and Adult Rat Hearts

  • Ramesh, Venkat;Kresch, Mitchell J.;Park, Woo-Jin;Kim, Do-Han
    • BMB Reports
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    • 제34권6호
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    • pp.573-577
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    • 2001
  • The mammalian heart is known to undergo significant mechanical changes during fetal and neonatal development. The objective of this study was to define the ontogeny of the ryanodine receptor/$Ca^{2+}$ release channel and SERCA that play the major roles in excitation-contraction coupling. Whole ventricular homogenates of fetal (F) (19 and 22 days in gestation), postnatal (N) (1 and 7 days postnatal), and adult (A) (5 weeks postnatal) Sprague-Dawley rat hearts were used to study [$^3H$]ryanodine binding and oxalate-supported $^{45}Ca^{2+}$ uptake. For the ryanodine receptor, the major findings were: (1) The ryanodine receptor density, as determined by maximal [$^3H$]ryanodine binding ($B_{max}$), increased 3 fold between the F22 and A periods ($0.26{\pm}0.1$ vs. $0.73{\pm}0.07$ pmoles/mg protein, p<0.01), whereas there was no significant change during the F22 and N1 development phases ($0.26{\pm}0.1$ vs. $0.34{\pm}0.01$). (2) Affinity of the ryanodine receptor to ryanodine did not significantly change, as suggested by the lack of change in the $K_d$ during the development and maturation. For SERCA, changes started early with an increased rate of $Ca^{2+}$ uptake in the fetal periods (F19: $8.1{\pm}1.1$ vs. F22: $19.3{\pm}2.2$ nmoles/g protein/min; p<0.05) and peaked by 7 days (N7) of the postnatal age ($34.9{\pm}2.1$). Thus, we conclude that the quantitative changes occur in the ryanodine receptor during myocardial development. Also, the maturation of the $Ca^{2+}$ uptake appears to start earlier than that of the $Ca^{2+}$ release.

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감초가 면역반응에 미치는 영향 (I) - 50% 메탄올 엑스의 면역조절작용 - (Effect of Glycyrrhizae Radix on the Immune Responses(I) - Immuno-regulatory Action of 50% Methanol Extract -)

  • 한종현;오찬호;은재순
    • 약학회지
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    • 제35권3호
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    • pp.154-164
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    • 1991
  • These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

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흰쥐 肝미토콘드리아의 非代謝依存性 칼슘 結合에 관한 연구 (Studies on the Metabolism-Independent Calium Binding of the Rat Liver Mitochondria)

  • Kang, Shin-Sung;Ha, Doo-Bong
    • 한국동물학회지
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    • 제13권3호
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    • pp.85-93
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    • 1970
  • 미토콘드리아의 Ca 결합은 물질대사에 의존하지 않는 初期 結合과 에너지 소비를 수반하는 膜透過의 二段階로 일어 난다는 설이 있다. 본 실험에서는 이를 확인키 위하여 흰쥐의 肝에 서 抽出한 미토콘드리아를 $^45 CaCl_2$를 함유한 sucrose-tris chloride 용액에서 incubate 시키면 서 Ca 흡수, 산소소비 및 ATPase 活性을 측정하였다. 미토콘드리아의 Ca 결합량은 온도의 영향을 거의 받지 않으며, succinate 나 ATP의 존재에 의해서도 증가하지 않는다. 반면 산소소비량은 succinate의 존재에 의하여 현저하게 증가되며 또 온도의 상승에 따라 증가된다. ATPase 活性도 온도의 상승에 따라 증가한다. 산소소비량과 Ca의 결합량이 비례하지 않는 것으로 보아 미토콘드리아의 Ca 초기 결합은 물질 대사에 의존하지 않는 것으로 판단된다. 미토콘드리아의 ATPase 活性은 DNP 의 존재에 의하여 증가된다.

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Phorbol Ester-Induced Periodic Contraction in Isolated Rabbit Jugular Vein

  • Ryu, Jae-Cheol;Jung, Dong-Keun;Lee, Sang-Ho
    • The Korean Journal of Physiology
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    • 제29권2호
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    • pp.225-232
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    • 1995
  • The present study was conducted to evaluate the effect of phorbol 12,13-dibutyrate (PDBu) on the contraction of rabbit jugular vein in vitro. PDBu concentrations of greater than 10 nM induced a periodic contraction which was composed of rapid contraction, plateau and slow relaxation. The frequency of periodic contraction increased as PDBu concentration increased. The PDBu-induced contraction was inhibited by staurosporine (100 nM), it was not changed by tetrodotoxin $(1\;{\mu}M).$ In $Ca^{2+}$-free medium, PDBu induced a sustaining contraction, but not periodic contraction. Addition of $Ca^{2+}$ to medium evoked periodic contraction which was inhibited by nifedipine, PDBu concentrations of greater than $0.1\;{\mu}M$ increased ^{45}Ca^{2+}$ uptake without changing $^{45}Ca^{2+}$ efflux. Charybdotoxin and apamin, $Ca^{2+}$-activated K^{+}$ channel blockers, did not affect the PDBu-induced periodic contraction, whereas tetraethylammonium (TEA) abolished the periodicity. Pinacidil $(10\;{\mu}M).$, a potassium channel activator, blocked PDBu induced periodic contraction, which was recovered by glybenclamide $(10\;{\mu}M).$. In high potassium solution, PDBu did not produce the periodic contraction. These results suggest that the PDBu-induced periodicity of contraction is modulated by voltage dependent $Ca^{2+}$ channel and ATP-sensitive $K^{+}$ channel.

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Identification of novel $Ca^{2+}$ binding proteins in junctional sarcoplasmic reticulum of rabbit skeletal muscle

  • Jung, Dai-Hyun;Mo, Sang-Hyun;Kim, Do-Han
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2002년도 제9회 학술 발표회 프로그램과 논문초록
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    • pp.56-56
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    • 2002
  • Muscle contraction and relaxation are regulated by the sarcoplasmic reticulum (SR)-mediated $Ca^{2+}$ release and $Ca^{2+}$ uptake. The SR functions are closely related with the proteins residing in the SR such as ryanodine receptor, $Ca^{2+}$-ATpase, calsequestrin, triadin and junctin. In an effort to further identify important functional SR proteins, experiments of sucrose-density gradient of SR fractionation, concanavalin A treatment, 2D gel electrophoresis, $^{45}$ Ca$^{2+}$ overlay, Strains-all staining, and peptide finger printing (PFP) were carried out.(omitted)d)

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감초가 면역반응에 미치는 영향(II) - Glycyrrhizin 및 Glycyrrhetinic acid의 면역조절작용 - (Effect of Glycyrrhizae Radix on the Immune Responses(II) - Immuno-regulatory Action of Glycyrrhizin and Glycyrrhetinic Acid -)

  • 한종현;오찬호;은재순
    • 약학회지
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    • 제35권3호
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    • pp.174-181
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    • 1991
  • These experiments were conducted to investigate the effects of glycyrrhizin(GL) and glycyrrhetinic acid(GA) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [sup 3/H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}$Ca$^{2+}$ to P388D$_{1}$, cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GL(10$^{-3}$M) and GA(10$^{-4}$M). Histamine production in mouse spleen cell culture was significantly increased by the addition of 0.25 $\mu\textrm{g}$/ml of Con A, after 48 hour incubation. Con A dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 .mu.g/ml of Con A. The effects of GL on histamine contents and T-lymphocyte proliferation were significantly decreased at high dose (10$^{-5}$M), while IL-1 activity was remarkably suppressed by 10$^{-8}$~10$^{-4}$M of GL. $Ca^{2+}$ uptake was not changed, but antibody production was increased by GL(10 mg/kg). GA inhibited histamine contents at 10$^{-9}$~10$^{-7}$ and depressed Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-7}$~10$^{-5}$M of GA, but increased suboptimal dose (Con A 0.1 $\mu\textrm{g}$/ml) at 10$^{-9}$~10$^{-7}$M of GA. IL-1 activity was suppressed by 10$^{-8}$~10$^{-4}$M of GA and $Ca^{2+}$ uptake was enhanced by 10$^{-9}$~10$^{-6}$ of GA, but antibody production was not changed by GA. From the above results, it is suggested that GL and GA have immuno-regulatory action. GL decreased cell-mediated immune response, and increased humoral immune response at high dose. On the other hand, low dose of GA enhanced cell-mediated immune response, while high doses of GA decreased humoral immune reaction.

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Molecular Properties of Excitation-Contraction Coupling Proteins in Infant and Adult Human Heart Tissues

  • Jung, Dai Hyun;Lee, Cheol Joo;Suh, Chang Kook;You, Hye Jin;Kim, Do Han
    • Molecules and Cells
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    • 제20권1호
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    • pp.51-56
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    • 2005
  • Excitation-contraction coupling (ECC) proteins in the human heart were characterized using human atrial tissues from different age groups. The samples were classified into one infant group (Group A: 0.2-7 years old) and three adult groups (Group B: 21-30; Group C: 41-49; Group D: 60-66). Whole homogenates (WH) of atrial tissues were assayed for ligand binding, $^{45}Ca^{2+}$ uptake and content of ECC proteins by Western blotting. Equilibrium [$^3H$]ryanodine binding to characterize the ryanodine receptor (RyR) of the sarcoplasmic reticulum (SR) showed that the maximal [$^3H$]ryanodine binding ($B_{max}$) to RyR was similar in all the age groups, but the dissociation constant ($k_d$) of ryanodine was higher in the infant group than the adult groups. Oxalate-supported $^{45}Ca^{2+}$ uptake into the SR, a function of the SR SERCA2a activity, was lower in the infant group than in the adult groups. Similarly, [$^3H$]PN200-110 binding, an index of dihydropyridine receptor (DHPR) density, was lower in the infant group. Expression of calsequestrin and triadin assessed by Western blotting was similar in the infant and adult groups, but junctin expression was considerably higher in the adult groups. These differences in key ECC proteins could underlie the different $Ca^{2+}$ handling properties and contractility of infant hearts.

사람적혈구막에서의 Calcium과 Suramin의 상호작용 (Interaction of Calcium with Suramin in Human Red Cell Preparation)

  • 강복순;강두희
    • The Korean Journal of Physiology
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    • 제10권1호
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    • pp.1-5
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    • 1976
  • The trypanocidal drug suramin, an impermeant polyanion, has been shown to be a powerful inhibitor of the calcium uptake and calcium-stimulated ATPase activity of sarcoplasmic reticulum (Fortes et al., 1974). In view of this finding, an attempt was made to investigate the effect of suramin on $Ca^{++}$ transport in resealed red cells and on $Ca^{++}$-activated ATPase in red blood cell membrane fragments (RBCMF). The results obtained are summarized as follows. 1. $Ca^{++}$ outflux from the resealed RBC was inhibited by suramin and the inhibitory action of suramin is proportional to the concentration of drug added inside the RBC preparation. When suramin is added both inside and outside the RBC preparation simultaneously, the magnitude of the inhibitory effect was more pronounced, suggesting that suramin inhibits both active $Ca^{++}-^{45}Ca$ exchange diffusion across the RBC membrane. 2. Suramin inhibits the $Ca^{++}$-activated ATPase of the RBCMF and the effect of inhibition by the drug was also concentration dependent. From the above results, it may be concluded that suramin inhibits $Ca^{++}$ transport across RBC membrane by inhibiting $Ca^{++}$-activated ATPase activity which has been known to be linked with active $Ca^{++}$ transport.

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성향정기산(星香正氣散)이 가토(家兎)의 경동맥(頸動脈) 평골근(平滑筋) 절편(切片)에서 t-Butyl Hydroperoxide 에 의한 지질과산화(脂質過酸化) 및 수축(收縮)에 미치는 영향(影響) (Effect of Sunghyangchungi-san (Xingxiangzhengqi-san) on Contraction and Lipid Peroxidation Induced by t-Butyl Hydroperoxide in Isolated Rabbit Carotid Artery)

  • 김영균;김종훈
    • 대한한의학회지
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    • 제20권3호
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    • pp.77-86
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    • 1999
  • This study was undertaken to evaluate the effect of Sunghyangchungi-san (SHCS) on the oxidant-induced contraction and lipid peroxidation in rabbit carotid artery. Vascular rings isolated from rabbit carotid artery were exposed to t-butyl hydroperoxide (t-BHP), an extrinsic oxidant, and the effect of SHCS on the changes of vascular tension and lipid peroxidation induced by t-BHP was determined. t- BHP induced a slowly developing and sustained contraction of the arterial rings. SHCS effectively relaxed the arterial rings that were pre-contracted by t-BHP. The responses to SHCS were partially dose-dependent at concentrations lower than 0.5 mg/ml. When SHCS was applied prior to the exposure to t-BHP, it inhibited the t-BHP-induced contraction as well. t- BHP increased lipid peroxidation in a dose-dependent manner. SHCS as well as well-known anti-oxidants GSH and DPPD reduced significantly lipid peroxidation induced by t-BHP. SHCS partially blocked the increase in $^{45}Ca$ uptake induced by t-BHP. In contrast to SHCS, anti-oxidants GSH and DPPD failed to inhibit significantly the t- BHP-induced contraction or $^{45}Ca$ uptake. From the above results, it is suggested that SHCS relaxed t-BHP-induced contraction of rabbit carotid artery independently of its anti-oxidant action, and inhibition of $Ca^{2+}$ influx may contribute to the underlying mechanism.

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