• Title/Summary/Keyword: $\Ca^{2+}$-ATPase

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Freeze Denaturation of Squid Actomyosin

  • Lee Kang-Ho;Ryu Hong-Soo;Cho Young-Je;Jung Byung-Chun;Hong Byung-Il;Je Yoi-Kwon;Lee Goon-Ja
    • Fisheries and Aquatic Sciences
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    • v.2 no.1
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    • pp.12-16
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    • 1999
  • Denaturation of actomyosin from the obliquely striated mantle muscle of squids (Todarodes pacificus) was studied by measuring the changes in $Ca^{2+}$-ATPase activity, relative viscosity, and solubility during frozen storage at three different temperature zones of maximum ice crystal formation $(-3^{\circ}C,\;-\;-5^{\circ}C)$, the eutectic point $(-11^{\circ}C)$, and $-20^{\circ}C$. The logarithms of $Ca^{2+}$-ATPase activity, relative viscosity and solubility of the actomyosin solutions (0.6 M KCl) and suspensions (0.05 M KCl) tended to decrease during frozen storage. The denaturation of squid actomyosin at the zone of maximum ice crystal formation significantly differed by only two degree of temperature difference between $-3^{\circ}C$ and $-5^{\circ}C$, and it (0.05 M KCl) at $-3^{\circ}C$ was less than those of other temperature. The denaturation at $-11^{\circ}C$ was more rapid than at $-5^{\circ}C$. The logarithms of $Ca^{2+}$ -ATPase activity, relative viscosity, and solubility were changed slower in the suspensions (0.05 M KCl) than the solutions (0.6 M KCl) at all experimental temperatures.

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$[^3H]$ Ouabain Binding and Effect of Ouabain on $^{45}Ca^{2+}$-Uptake in Rat Cardiac Myocytes (쥐 심근 세포의 $[^3H]$ Ouabain 결합과 $^{45}Ca^{2+}}$섭취에 미치는 Ouabain의 영향)

  • 이신웅;김영희;진갑덕
    • YAKHAK HOEJI
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    • v.28 no.3
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    • pp.129-138
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    • 1984
  • Specific [$^{3}H$] ouabain binding and $Ca^{2+}$ -uptake were measured to elucidate the role of high affinity [$^{3}H$] ouabain binding site in rat cardiac myocytes which contain 65% of rod cells. High affinity [$^{3}$H] ouabain binding site, which is about 3% of total pump sites, with apparent dissociation constant ($K_{D}$) of $1.1{\times}10^{-7}M$ and maximum binding site concentration (Bmax) of 1.2 pmol/mg protein ($1.754{\times}10^{5}cells$) were identified. At the concentration of $10^{-7}M$ to $10^{-4}M$, ouabain produced concentration dependent increase in $Ca^{2+}$-uptake of myocytes. The effect of ouabain on $Ca^{2+}$-uptake was not effected by membrane depolarization (elevated K+ in incubation medium) or verapamil. These results suggest that in rat ventricular myocytes the ouabain receptor complex to high affinity site may increase Na+ - $Ca^{2+}$ exchange across the sarcolemmal membrane by inhibition of Na+, K+ - ATPase.

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Effects of Inositol 1,4,5-triphosphate on Osteoclast Differentiation in RANKL-induced Osteoclastogenesis

  • Son, A-Ran;Kim, Min-Seuk;Jo, Hae;Byun, Hae-Mi;Shin, Dong-Min
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.1
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    • pp.31-36
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    • 2012
  • The receptor activator of NF-${\kappa}B$ ligand (RANKL) signal is an activator of tumor necrosis factor receptor-associated factor 6 (TRAF6), which leads to the activation of NF-${\kappa}B$ and other signal transduction pathways essential for osteoclastogenesis, such as $Ca^{2+}$ signaling. However, the intracellular levels of inositol 1,4,5-trisphosphate ($IP_3$) and $IP_3$-mediated cellular function of RANKL during osteoclastogenesis are not known. In the present study, we determined the levels of $IP_3$ and evaluated $IP_3$-mediated osteoclast differentiation and osteoclast activity by RANKL treatment of mouse leukemic macrophage cells (RAW 264.7) and mouse bone marrow-derived monocyte/macrophage precursor cells (BMMs). During osteoclastogenesis, the expression levels of $Ca^{2+}$ signaling proteins such as $IP_3$ receptors ($IP_3Rs$), plasma membrane $Ca^{2+}$ ATPase, and sarco/endoplasmic reticulum $Ca^{2+}$ ATPase type2 did not change by RANKL treatment for up to 6 days in both cell types. At 24 h after RANKL treatment, a higher steady-state level of $IP_3$ was observed in RAW264.7 cells transfected with green fluorescent protein (GFP)-tagged pleckstrin homology (PH) domains of phospholipase C (PLC) ${\delta}$, a probe specifically detecting intracellular $IP_3$ levels. In BMMs, the inhibition of PLC with U73122 [a specific inhibitor of phospholipase C (PLC)[ and of $IP_3Rs$ with 2-aminoethoxydiphenyl borate (2APB; a non-specific inhibitor of $IP_3Rs$) inhibited the generation of RANKL-induced multinucleated cells and decreased the bone-resorption rate in dentin slice, respectively. These results suggest that intracellular $IP_3$ levels and the $IP_3$-mediated signaling pathway play an important role in RANKL-induced osteoclastogenesis.

Changes in Morphologic and Enzymatic Properties of Beef Myofibrillar Protein by Storage Tmeperature (저장온도에 따른 쇠고기 근원섬유의 형태적, 효소적 성질 변화)

  • 정인철
    • The Korean Journal of Food And Nutrition
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    • v.10 no.4
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    • pp.468-474
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    • 1997
  • This study was designed to investigate the changes in meat quality of beef shank, rib and loin during storage at 8$^{\circ}C$. The shear force value(SFV) of beef shank and loin decreased significantly after 6days storage, beef loin was no significant difference during storage. The SFV in early storage period was high in the order of beef rib, loin and shank, but the SFV of beef rib and loin was similar in course of storage period. The Myofibrillar fragmentation index(MFI) of beef shank increased significantly after 6 days storage, but beef rib and loin early storage was high in the order of beef rib, loin and shank. The actomyosin extractability after 3days storage increased in all parts of beef, but beef loin decreased after 6 days storage. In case of Mg2+-ATPase activity of actomyosin, beef shank increased to 3 days storage, and this reached the level of 0 day after 6days. The MG2+-ATPase activity of beef rib and loin was similar, but beef rib in early storage was higher than beef loin. The Ca2+-TPase activity of beef shank increased to 3 days and decreased after 6 days storage, beef rib was not different during storage and beef loin decreased slightly during storage.

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A Comparative Study on Spermatozoan Maturation in the Hibernating Animals (동면동물의 정자성숙과정에 대한 비교 연구)

  • Jae-Ho Chang;Yung-Keun Oh;In-Ho Choi;Noh-Pal Jung;Hyung-Cheul Shin
    • Biomedical Science Letters
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    • v.3 no.1
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    • pp.1-9
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    • 1997
  • Acrosome reaction usually occures just before fertilization in most mammals, and it has been known that $Ca^{2+}$ plays an important role in the acrosome reaction and albumin also known as a critical factor for spermatozoan activities. The present study has been designed in order to observe maturing processes of the spermatozoa occurred in the ductus epididymidis and to clarify the relationships of $Ca^{2+}$ concentrations with those processes, and to compare the enzymatic activities of ATPase and the lactate dehydrogenase of the spermatozoa in accordance with time before and after the spermatozoan maturation. From the results, we can confirm that most of the bat spermatozoa come to maturity within the epididymal cauda and may pass through capacitation outside the cauda. However it is expected to be studied that the fluctuation of spermatogenic activity depending on temperature changes and their relationships with the ductus epididymidis and their mutual influences.

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$Ca^{2+}$ Effect on Conversion of Exogenous 1-Aminocyclopropane-1-Carboxylic Acid to Ethylene in Vigna radiata Protoplasts

  • Seung-Eun Oh
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.271-276
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    • 1994
  • The possibility that 1-aminocyclopropane-1-carboxylic acid (ACC)-uptake may be dependent on the H+-gradient established across the plsma membrane was tested in protoplasts isolated from 2.5 day old mungbean hypocotyls. The ACC-induced ethylene production was inhibited when the H+-gradient was collapsed by the treatment with carbonycyamide-p-trifluro-methoxy-phenylhydrazone (FCCP). Moreover, the treatment with o-vanadate, a specific inhibitor of plasma membrane H+-ATPase, caused the inhibition of ethylene production. The ACC-induced ethylene production was inhibited by the treatemnt with verapamil (Ca2+-channel blocker), or ethylene glycol-bis($\beta$-aminoethyl ether) N, N, N', N'-tetraacetic acid (EGTA) (Ca2+-chelator). In contrast, the ehtylene production was stimulated by the application of A23187 (Ca2+ ionophore). The inhibitory effect of EGTA in the ethylene producton was magnified in the presence of A23187. From these results, we suggest that the external Ca2+ influx to the cytosol resulted in the stimulatin of ACC oxidase activity after ACC-uptake resulting from a H+-gradient across the plasma membrane.

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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
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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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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Role of Regulators of G-Protein Signaling 4 in $Ca^{2+}$ Signaling in Mouse Pancreatic Acinar Cells

  • Park, Soon-Hong;Lee, Syng-Ill;Shin, Dong-Min
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.6
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    • pp.383-388
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    • 2011
  • Regulators of G-protein signaling (RGS) proteins are regulators of $Ca^{2+}$ signaling that accelerate the GTPase activity of the G-protein ${\alpha}$ -subunit. RGS1, RGS2, RGS4, and RGS16 are expressed in the pancreas, and RGS2 regulates G-protein coupled receptor (GPCR)-induced $Ca^{2+}$ oscillations. However, the role of RGS4 in $Ca^{2+}$ signaling in pancreatic acinar cells is unknown. In this study, we investigated the mechanism of GPCR-induced $Ca^{2+}$ signaling in pancreatic acinar cells derived from $RGS4^{-/-}$ mice. $RGS4^{-/-}$ acinar cells showed an enhanced stimulus intensity response to a muscarinic receptor agonist in pancreatic acinar cells. Moreover, deletion of RGS4 increased the frequency of $Ca^{2+}$ oscillations. $RGS4^{-/-}$ cells also showed increased expression of sarco/endoplasmic reticulum $Ca^{2+}$ ATPase type 2. However, there were no significant alterations, such as $Ca^{2+}$ signaling in treated high dose of agonist and its related amylase secretion activity, in acinar cells from $RGS4^{-/-}$ mice. These results indicate that RGS4 protein regulates $Ca^{2+}$ signaling in mouse pancreatic acinar cells.

육질향상처리가 생선횟감용 어류 근육의 물리ㆍ화학적 변화에 미치는 영향 연구 5. 저장 중 넙치육의 ATPase활성 변화

  • 이기봉;심길보;조민성;김태진;조영제
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2002.10a
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    • pp.118-119
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    • 2002
  • 근육이 수축하면 근원섬유의 sarcomere (근절)의 길이가 경직의 진행과 함께 짧아지는 것을 볼 수 있다. 즉, thick filament (myosin filament)와 thin filament (actin filament) 사이에서로 미끌어져 들어가는 현상이 일어나게 되는 것이다. 골격근은 보통 때는 이완상태에 있으나 필요할 때 신경자극에 의해서 수축을 하게 된다. 이러한 수축↔이완의 전환은 세포내의 $Ca^{2+}$ 농도의 조절을 통해서 제어되며, 이완시의 세포질 내의 $Ca^{2+}$ 농도는 정도의 낮은 상태로 유지된다. (중략)

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