• Title/Summary/Keyword: PLD2

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Purification and Characterization of Proteins Inhibiting Phospholipase D Activity from Flounder (Paralichthys olivaceus) Brain (넙치 (Paralichthys olivaceus) 뇌로부터 phospholipase D 활성 억제 단백질의 정제 및 특성 규명)

  • SEO Jung-Soo;KIM Eun-Hi;HWAWG Eun-Young;KIM Nam Deuk;KIM Dong Sun;LEE Hyung-Ho;CHUNG Joon-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.4
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    • pp.370-377
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    • 2001
  • Flounder brain cytosol contains protein inhibitors that markedly inhibit the activity of partially purified brain membrane phospholipase D (PLD) which is dependent on phosphatidylinositol 4,5-bisphosphate ($PIP_2$) but insensitive to ADP-ribosylation factor (ARF), The PLD inhibitors have been enriched through several chromatographic steps and characterized with respect to size and mechanism of inhibition. Sequential chromatography of the brain cytosol yielded six inhibitor fractions, Two (IIA and IIB) of six inhibitor fractions showed the $PIP_2$-phosphatase activities. IIA was identified as synaptojanin, a nerve terminal protein that has known to be a member of the inositolpolyphosphate 5-phosphatase family, by immunoblot analysis. IIB showed an apparent molecular mass of 158 kDa by Superose 12 gel filtration chromatography and was immunologically distinct from synaptojanin. IIB hydrolyzed $PIP_2$, yielding only phosphatidylinositol phosphate (PIP) as product, suggesting that IIB hydrolyzes only one phosphate from either the 4- or 5-position of PI (4,5)$P_2$. These studies demonstrate that the existence of multiple $PIP_2$-phosphatases have been implicated in the negative regulation of $PIP_2$-dependent PLD activity within flounder brain.

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Effect of Alcohols Toward the Transphosphatidylation Activity in Phospholipase D Catalyzed Reaction (포스포리파제 D 촉매반응에서 포스파티딜 전달반응 활성에 미치는 알코올의 영향)

  • Koh, Eun-Hie
    • Journal of the Korean Chemical Society
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    • v.54 no.2
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    • pp.208-214
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    • 2010
  • In the presence of alcohol, phospholipase D (PLD) is known to perform transphosphatidylation activity, during which the overall reaction rate of PLD increased. To elucidate the reaction mechanism of transphosphatidylation further, we investigated rate constants of transphosphatidylation reaction of the purified ${\alpha}$-type PLD from cabbage in the presence of various alcohols. The second-oder rate constants of PLD transphosphatidylation showed a large increase with the primary alcohols examined as expected. In the case of butanol we observed the second-oder rate constant of $33.33{\pm}1.33M^{-1}sec^{-1}$. This second-order rate constant of transphosphatidylation was as 400 times greater as the second-order hydrolysis rate constant of $0.078M^{-1}sec^{-1}$ which was adjusted for the water concentration. A linear free energy relationship between the $pK_a$ of alcohol and transphosphatidylation rate gives a Br${\o}$nsted slope of ${\beta}_{nu}$ = 0.12 ${\pm}$ 0.03. This small ${\beta}_{nu}$ value implicates that the transition state of break down of phosphatidyl-enzyme intermediate (E-P) is likely dissociative. Finally, a reaction mechanism of cabbage PLD is suggested on the basis of our results presented here and the histidine residue known to be located in the active site of cabbage PLD.

Fabrication of YBCO coated conductors by PLD continuous reel-to-reel processing (PLD 연속 공정을 통한 YBCO coated conductor 제조)

  • ;;Donggqi Shi
    • Proceedings of the Korea Institute of Applied Superconductivity and Cryogenics Conference
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    • 2003.10a
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    • pp.150-152
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    • 2003
  • YBa$_2$Cu$_3$O$_{7-{\delta}}$(YBCO) coated conductors were deposited by pulsed laser deposition (PLD) on short buffered substrate in continuous PLD reel-to-reel system. The oxide multilayer buffered substrate of architectures of CeO$_2$/YSZ/Y$_2$O$_3$was fabricated by PLD at steady status. The degree of texture of each layer was investigated using X-ray diffraction including $\theta$-2$\theta$ scans, $\omega$-scans and $\Phi$-scans analysis. Their surface morphology was observed by scanning electron microscopy (SEM) The FWHM of the X-ray $\omega$-scans and $\Phi$-scans indicated that YBCO and buffer layers closely replicate the in-plane and out-of-plane texture of metal tape. Critical current at 77K self-field of 19A, critical temperature of 86K, and current density of 2MA/$\textrm{cm}^2$ were measured. The film also exhibits a homogeneous and dense surface morphology.e morphology.

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Screenin of Phospholipase D Producing Actinomycetes (방선균으로부터 Phospholipase D 생산균주의 탐색)

  • 손동화;심재용;윤석후
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.333-339
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    • 1994
  • In order to screen microorganisms producing phospholipase D (PLD) [EC 3.1.4.4], culture broths of about 900 strains of soil bacteria were subjected to examine for the PLD activity. When the hydrolytic activity of PLD (H-activity) in the supernatant was determined, 64 strains produced PLD more than 0.3 unit/ml and all of them were actinomycetes. Among 26 culture broths tested, 6 ones had transphosphatidylation activity (T-activity) of 30~68%. When the strains except one were cultivated on 3 different media at 30$\circ$C for 3 days under aerobic condition, strain # 1090 on medium B (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, CaCO$_{3}$ 0.4%, and pH 7.2) produced PLD with much higher H- and T-activity, which were 8.3 units/ml and 76.3%, respectively. Subsequently, time course of PLD production of the strain # 1090 during cultivation with aeration of 1 v/v/m and agitation of 400 rpm at 30$\circ$C for 5 days on medium B in jar fermentor was investigated. H-activty of PLD reached almost maximum (about 9 units/ml) after 32 hours and maximal T-activity was found to be about 80%.

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Preparation of $NbS_2$ thin film using PLD method (PLD 장치를 이용한 $NbS_2$ 박막의 제작)

  • Park, Jong-Man;Lee, Hea-Yeon;Jeong, Jung-Hyun
    • Journal of Sensor Science and Technology
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    • v.7 no.5
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    • pp.372-376
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    • 1998
  • We developed a pulsed laser deposition(PLD) apparatus for depositing various thin films. In this study, the formation of $NbS_2$ thin film was performed in the vacuum chamber by PLD method. $Al_2O_3$(012) and Si(111) were used as the substrates. In order to investigate the growth conditions of a high crystalline $NbS_2$ thin film, the S/Nb composition ratio was varied from 2.0 to 5.25 and the substrate temperature was varied from the room temperature to $600^{\circ}C$. From the result of X-ray diffraction studies of the prepared $NbS_2$ thin films, it was reported that the $NbS_2$, thin film showed a good crystallinity at substrate temperature $600^{\circ}C$ and with S/Nb composition ratio 4.0 on $Al_2O_3$(012) but did not on Si(111). The films exhibited c-axis orientation.

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펄스레이저를 이용한 박막 증착

  • 김호성
    • 전기의세계
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    • v.42 no.6
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    • pp.46-51
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    • 1993
  • 본 특집에서는 PLD장착 및 증착 방법을 2장에서는 소개하고 3장에서는 증착원리 및 특징을 4장에서는 PLD의 응용분야를 그리고 5장에서는 결론 및 향후 연구방향에 대하여 기술하였다.

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Epigallocatechin Gallate Activates Phospholipase D in Glioma Cells (교세포에서 Epigallocatechin Gallate에 의한 포스포리파제 D의 활성화)

  • Kim, Shi-Yeon;Kim, Joonmo;Min, Do-Sik
    • Journal of Life Science
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    • v.13 no.6
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    • pp.924-932
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    • 2003
  • Epigallocatechin-3 Gallate (EGCG), a major constituent of green tea, has attracted increasing interest because of its many reported health benefits. Here we demonstrate for the first time that EGCG stimulates phospholipase D (PLD) activity in U87 human astroglioma cells. EGCG-induced PLD activation was abolished by the phospholipase C (PLC) inhibitor and a lipase inactive PLC-\gama1$ mutant, and was dependent on intracellular $Ca^{ 2+}$, and possibly involved $Ca^{ 2+}$ calmodulin-dependent protein kinase II (CaM kinase II). Interestingly, EGCG induced translocation of PLC-\gama1$ from the cytosol to the membrane and PLC-\gama1$interaction with PLD1. Taken together, these results demonstrate for the first time that in human astroglioma cells, EGCG regulates PLD activity via a signaling pathway involving a PLC-\gama1$ (inositol 1,4,5-trisphosphate-$Ca^{ 2+}$)-CaM kinase II-PLD pathway.

The Effects of Single Component of Ginsenosides on the Mechanism of Mediator Release in the Allergic Hypersensitivity (인삼 사포닌 단일물질이 알러지 과민반응의 매개체 유리기전에 미치는 영향)

  • Ro, Jai-Youl;Kim, Kyung-Hwan
    • The Korean Journal of Pharmacology
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    • v.30 no.2
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    • pp.243-254
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    • 1994
  • Inflammatory diseases, allergic and asthmatic disorders are caused by the mediator release from the activation of the phospholipase C (PLC), phospholipase D (PLD), methyltransferase or adenylate cyclase etc. during IgG or IgE cross-linking of high affinity receptors on mast cells or basophil surface. One important enzyme activated after IgG or IgE receptor cross-linking is PLD, the enzyme which converts phosphatidylcholine (PC) to phosphatidic acid (PA). Under the hypothesis that these may be some differences in mediator release according to the difference in PLD activity, we attempted to confirm the ginseng saponin effects on the PLD activity. We examined the PLD activity during the passively sensitized mast cell activation in the presence of single component of ginsenosides $(Rc,\;Rg_1,\;Rg_2,\;Rg_3)$. We also measured the amount of mediators (histamine and leukotrienes) released by stimulating with ovalbumin (OA) or calcium ionophore (CaI), Guinea Pig lung mast cells were purified using enzyme digestion, count current elutriation, and discontinuous Percoll density gradient. In purified mast cells prelabeled with $[^3H]$ arachidonic acid or $[^3H]$ palmitic acid, PLD activity was assessed more directly by the production of labeled PEt by PLD-mediated transphosphatidylation in the presence of ethanol. Histanine release was determined by Spectrophotofluorometry, and leukotrienes by radioimmunoassay. The PLD activity during the passively sensitized mast cell activation is increased up to $3{\sim}5times$. The PLD activity during the passively sensitized mast cell activation in the presence of all ginsenosides is decreased up to $4{\sim}11$ times. $Rg_l\;and\;Rg_2$ ginsenoside pretreatment decreased histamine and leukotrienes by 50% in the OA-induced or by 40% in the Cal-induced mast cell after passively sensitization. Rc pretreatment poorly decreased histamine but leukotrienes decreased by 70% in the OA-induced or by 35% in the Cal-induced mast cell. $Rg_3$ ginsenoside pretreatment increased histamine release without challenging OA or Cal but leukotrienes decreased. These observations indicate that single unit of ginsenosldes may be an important contributor to inhibit the release of histamine and leukotrienes in the guinea pig lung mast cells, that inhibits the PLD-mediated formation of DAG evoked by mast cell activation.

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Characteristics of $Cu_2ZnSnSe_4$ Thin Film Solar Absorber Prepared by PLD using Solid Target (광흡수층 적용을 위한 PLD용 $Cu_2ZnSnSe_4$ 타겟 제조와 증착 박막의 특성)

  • Jung, Woon-hwa;Rachmat, Adhi Wibowo;Kim, Kyoo-ho
    • 한국신재생에너지학회:학술대회논문집
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    • 2009.06a
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    • pp.130-133
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    • 2009
  • $Cu_2ZnSnSe_4$(CZTSe) is one of the promising materials for the solar cell due to its abundant availability in the nature. In this study, we report the fabrication of CZTSe thin film by Pulsed Laser Deposition(PLD) method using quaternary compound target on sodalime glass substrate. The quaternary CZTSe compound target was synthesized by solid state reaction method using elemental powders of Cu, Zn, Sn and Se. Powders were milled in high purity ethanol using zirconia ball with mixed size of 1 and 3 mm at the same proportions for 72 hours milling time. The structural, chemical and mechanical properties of the synthesized CZTSe powders were investigated prior to the deposition process. The CZTSe compound powder, and $500^{\circ}C$ of sintering temperature shows the best properties for PLD target. Results show that the as-deposited CZTSe thin films with the precursors by PLD have a composition near-stoichiometric.

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Development of a PLD heater for continuous deposition and growth of superconducting layer

  • Jeongtae Kim;Insung Park;Gwantae Kim;Taekyu Kim;Hongsoo Ha
    • Progress in Superconductivity and Cryogenics
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    • v.25 no.2
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    • pp.14-18
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    • 2023
  • Superconducting layers deposited on the metal substrate using the pulsed laser deposition process (PLD) play a crucial role in exploring new applications of superconducting wires and enhancing the performance of superconducting devices. In order to improve the superconducting property and increase the throughput of superconducting wire fabricated by pulsed laser deposition, high temperature heating device is needed that provides high temperature stability and strong durability in high oxygen partial pressure environments while minimizing performance degradation caused by surface contamination. In this study, new heating device have been developed for PLD process that deposit and growth the superconducting material continuously on substrate using reel-to-reel transportation apparatus. New heating device is designed and fabricated using iron-chromium-aluminum wire and alumina tube as a heating element and sheath materials, respectively. Heating temperature of the heater was reached over 850 ℃ under 700 mTorr of oxygen partial pressure and is kept for 5 hours. The experimental results confirm the effectiveness of the developed heating device system in maintaining a stable and consistent temperature in PLD. These research findings make significant contributions to the exploration of new applications for superconducting materials and the enhancement of superconducting device performance.