• Title/Summary/Keyword: $cPLA_2$

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Identification and Characterization of a Novel Cytosolic Phospholipase from Bovine Red Blood Cells (소 적혈구로부터 새로운 형태의 세포질 포스포리파제 의 동정 및 특성규명)

  • 신혜숙;김하동;장동훈;전형준;유충규
    • YAKHAK HOEJI
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    • v.45 no.4
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    • pp.407-412
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    • 2001
  • A $Ca^{2+}$_dependent PLA$_2$activity termed rPLA$_2$, was detected in the cytosol of bovine red blood cells (RBCs). The rPLA$_2$was characterized as a similar form to Group IV cPLA$_2$, but different in several column chromatographic profiles including an anion exchange column. To examine whether this rPLA$_2$is different from the well characterized cPLA$_2$, a quinone derivative, BJ50, was developed and tested for the inhibitory effect on the two PLA$_2$enzymes. The rPLA$_2$activity was inhibited by a quinone derivative (BJ50) with ID$_{50}$ of 20 $\mu$M., but had no effect on the cPLA$_2$, suggesting that rPLA$_2$may be a novel type of cytosolic PLA$_2$in RBCs.s.s.

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Glutamine Inhibits TNF-α-induced Cytosolic Phospholipase A2 Activation via Upregulation of MAPK Phosphatase-1

  • Yoon, So Young;Jeong, Soo-Yeon;Im, Suhn-Young
    • Biomedical Science Letters
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    • v.27 no.4
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    • pp.223-230
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    • 2021
  • Tumor necrosis factor alpha (TNF-α) is a principal regulator of inflammation and immunity. The proinflammatory properties of TNF-α can be attributed to its ability to activate the enzyme cytosolic phospholipase A2 (cPLA2), which generates potent inflammatory lipid mediators, eicosanoids. L-glutamine (Gln) plays physiologically important roles in various metabolic processes. We have reported that Gln has a potent anti-inflammatory activity via rapid upregulation of mitogen-activated protein kinases (MAPKs) phosphatase (MKP)-1, which preferentially dephosphorylates the key proinflammatory enzymes, p38 MAPK and cytosolic phospholipase A2 (cPLA2). In this study, we have investigated whether Gln could inhibit TNF-α-induced cPLA2 activation. Gln inhibited TNF-α-induced increases in cPLA2 phosphorylation in the lungs and blood levels of the cPLA2 metabolites, leukotrine B4 (LTB4) (lipoxygenase metabolite) and prostaglandin E2 (PGE2) (cyclooxygenase metabolite). TNF-α increased p38 and cPLA2 phosphorylation and blood levels of LTB4 and PGE2, which were blocked by the p38 inhibitor SB202190. Gln inhibited TNF-α-induced p38 and cPLA2 phosphorylation and production of the cPLA2 metabolites. Such inhibitory activity of Gln was no longer observed in MKP-1 small interfering RNA-pretreated animals. Our data indicate that Gln inhibited TNF-α-induced cPLA2 phosphorylation through MKP-1 induction/p38 inhibition, and suggest that the utility of Gln in inflammatory diseases in which TNF-α plays a major role in their pathogenesis.

Quality of Strawberry Packed with PLA Films (PLA 필름으로 포장한 딸기의 품질)

  • Park, Hyung-Woo;Lee, Sun-Ah;Kim, Sang-Hee;Jeon, Seung-Ho
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.17 no.2
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    • pp.43-46
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    • 2011
  • Effect of several PLA films on strawberry packaging was investigated. Three types of PLA film (non-perforated PLA1, perforated films of PLA2 and PLA3) were used for packing strawberry and the fruit quality was measured during 6 day storage at 10. For comparison, two types of OPP/PE film (non-perforated OPP1 and perforated OPP2) were used for strawberry packaging. Weight loss of all the treatments was below 0.12% after 6 day, seeming to be in tolerable range. PLA3 package showed the best retention in fruit firmness, PLA2 and PLA3 packages the least changes in acidity, and PLA2 package gave the least changes of soluble solid content and vitamin C content. There have been no differences in surface color among packages. Perforated PLA films could be effective for use as packaging of strawberry.

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Freshness of Soybean Sprout Packed with PLA Films (저탄소 친환경 PLA 필름으로 포장한 콩나물의 신선도)

  • Park, Hyung-Woo;Kim, Sang-Hee;Jang, Jong-Gun
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.17 no.1
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    • pp.1-5
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    • 2011
  • Soybean sprouts of 100 g were packed with commercial OPP/PE films and developed PLA films, and stored at $10^{\circ}C$ for 6 days. Effects of packaging film were investigated. The weight loss of OPP/PE film was higher than that of the PLA film. Total cell count of PLA2 and PLA3 packages was the lowest among 5 packages. Off-flavor was strong at OPP/PE, but that in PLA2 was very little. Vitamin C content of PLA2 was highest and preference test of PLA2 was also highest among 5 packages. Finally, the PLA packages for soybean sprouts was more suitable than OPP/PE.

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Morinda citrifolia Inhibits Both Cytosolic $Ca^{2+}$-dependent Phospholipase $A_2$ and Secretory $Ca^{2+}$-dependent Phospholipase $A_2$

  • Song, Ho-Sun;Park, Sung-Hun;Ko, Myoung-Soo;Jeong, Jae-Min;Sohn, Uy-Dong;Sim, Sang-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.3
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    • pp.163-167
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    • 2010
  • This study investigated the effects of the methanol extracts of Morinda citrifolia containing numerous anthraquinone and iridoid on phospholipase $A_2$ ($PLA_2$) isozyme. $PLA_2$ activity was measured using various $PLA_2$ substrates, including 10-pyrene phosphatidylcholine, 1-palmitoyl-2-[$^{14}C$]arachidonyl phosphatidylcholine ([$^{14}C$]AA-PC), and [$^3H$]arachidonic acid (AA). The methanol extracts suppressed melittin-induced [$_3H$]AA release in a concentration-dependent manner in RAW 264.7 cells, and inhibited $cPLA_2/sPLA_2$-induced hydrolysis of [$^{14}C$]AA-PC in a concentration- and time-dependent manner. A Dixon plot showed that the inhibition by methanol extracts on $cPLA_2$ and $sPLA_2$ appeared to be competitive with inhibition constants ($K_i$) of $3.7{\mu}g/ml$ and $12.6{\mu}g/ml$, respectively. These data suggest that methanol extracts of Morinda citrifolia inhibits both $Ca^{2+}$-dependent $PLA_2$ such as, $cPLA_2$ and $sPLA_2$. Therefore, Morinda citrifolia may possess anti-inflammatory activity secondary to $Ca^{2+}$-dependent $PLA_2$ inhibition.

Detection and Characterization of Two Novel Forms of A Cytosolic Phospholipase $A_2$-Activating Factor in Bovine Brain

  • Park, Jae-Sin;Jung, Kwang-Mook;Jung, Sung-Yun;Na, Doe-Sun;Kim, Dae-Kyong
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.187-187
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    • 1996
  • 세포질에 존재하는 100 kDa Phospholipase $A_2$(cPLA$_2$)는 인지질의 sn-2 위치의 에스테르결합을 가수분해함으로서 Prostaglandin과 Leukotriene등 Eicosanoids 생합성의 전구체인 아라키돈산과 Platelet activating factor(PAF)를 생합성하는 전구체를 동시에 생성시키는 효소로 염증과 세포손상등에 중요한 역할이 기대된다. 본 효소의 활성화 기전을 규명하고자 하는 최근의 활발한 연구에도 불구하고 불명확한 점이 많은 것이 현실이다. 특히 세포를 자극하였을 때 유리되는 아라키돈산의 증가율과 세포를 파괴한 후 조제한 가용성분획에서 측정한 활성의 증가율과는 많은 차이를 나타냈다. 이러한 결과부터 cPLA$_2$ 효소 자체를 활성화시키는 어떤 인자를 가정하였다. 최근, PLA$_2$의 또다른 형태인 14 kDa의 분비성 PLA$_2$의 in vitro 활성을 증가시키는 인자가 동정되어 그 생화학적 특성이 규명되고 있으나 이 인자는 cPLA$_2$의 활성에는 아무런 증가효과를 나타내지 않았다. 본 연구자들은 소의 뇌조직에서 cPLA$_2$의 활성을 증가시키는 인자를 발견하고 그의 생화학적인 특성을 규명하였다. 돼지 비장에서 정제한 cPLA$_2$를 사용하였으며 소의 뇌 조직의 가용성 분획으로부터 본 활성화 인자를 동정하였으며 그 활성분획을 양이온 크로마토그라피로서 Mono S EPLC와 Superose 12 Sepharose gel filtration 크로마토그라피를 이용하여 더욱 분리한 결과 약 70 kDa과 25 kDa에서 각각 용출되었다. 이렇게 부분정제한 활성은 췌장에서 분리한 group I과 흰주의 group I과 흰주의 혈소판에서 분리한 group II PLA$_2$에 대해서는 아무런 증가효과를 나타내지 않는 반면, cPLA$_2$의 활성만을 약 5배 증가시켰다. 본 활성은 cPLA$_2$ 효소량의 증가에 따라 활성의 증가효과가 정차 감소하므로 화학량적인 반응(Stoichiometric reaction)일 것으로 예상되었다.

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Cytosolic Phospholipase A2 Activity in Neutrophilic Oxidative Stress of Platelet-activating Factor-induced Acute Lung Injury (Platelet-activating Factor에 의한 급성폐손상에서 호중구성 산화성 스트레스에 관여하는 Cytosolic Phospholipase A2 활성도의 변화)

  • Kwon, Young Shik;Hyun, Dae Sung;Lee, Young Man
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.6
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    • pp.497-506
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    • 2007
  • Background: The present investigation was performed in rats and isolated human neutrophils in order to confirm the presumptive role of the positive feedback loop of cytosolic phospholipase $A_2$ ($cPLA_2$) activation by plateletactivating factor (PAF). Methods: The possible formation of the positive feedback loop of the $cPLA_2$ activation and neutrophilic respiratory burst was investigated in vivo and in vitro by measurement of the parameters denoting acute lung injury. In addition, morphological examinations and electron microscopic cytochemistry were performed for the detection of free radicals in the lung. Results: Five hours after intratracheal instillation of PAF ($5{\mu}g/rat$), the lung leak index, lung myeloperoxidase (MPO) activity, the number of neutrophils and the concentration of cytokine-induced neutrophil chemoattractant (CINC) in bronchoalveolar lavage fluid were increased by PAF as compared with those of control rats. The NBT assay and cytochrome-c reduction assay revealed an increased neutrophilic respiratory burst in isolated human neutrophils following exposure to PAF. Lung and neutrophilic $cPLA_2$ activity were increased following PAF exposure and exposure to hydrogen peroxide increased $cPLA_2$ activity in the lung. Histologically, inflammatory findings of the lung were observed after PAF treatment. Remarkably, as determined by $CeCl_3$ cytochemical electron microscopy, increased production of hydrogen peroxide was identified in the lung after PAF treatment. Conclusion: PAF mediates acute oxidative lung injury by the activation of $cPLA_2$, which may provoke the generation of free radicals in neutrophils.

Study on Isothermal Crystallization Characteristics of PLA Film by Adding APP as a Nucleation Agent (APP 핵제를 첨가한 PLA 필름의 등온결정화 특성에 관한 연구)

  • Kim, Gyu-Sun;Kim, Moon-Sun;Kim, Byung-Woo
    • Korean Chemical Engineering Research
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    • v.50 no.3
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    • pp.582-587
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    • 2012
  • In this paper, it was studied on the crystallization characteristics of PLA film by adding ammonium phosphate (APP) as a nucleation agent. Crystallinity and crystallite size of PLA film were determined by Scherrer equation. Crystallization rate constant of PLA film was calculated through Avrami equation. Film samples in the study were prepared by two steps. PLA films were prepared by adding 1, 5, and 10 wt%, respectively, at first and was secondly annealed at 130, 140, and $150^{\circ}C$. Crystallinity of pure PLA film was average 4.6% and those of PLA film with adding 1, 5, and 10 wt% APP were 12.2, 47.7, and 50.0%, respectively. Crystallite size of PLA film was average 28.0 nm and those of PLA film with adding 1, 5, and 10 wt% APP were 26.8, 24.0, and 19.0 nm, respectively. Crystallization rate constants of PLA film with 1 wt% APP were 2.12, 3.86, and 0.27 by annealing at 130, 140, and $150^{\circ}C$, respectively, where was higher than pure PLA film and those with adding 5 and 10 wt% APP, respectively.

Freshness of Paprika Packed with PLA Films (개발 PLA 필름으로 포장한 파프리카의 신선도)

  • Park, Hyung-Woo;Kim, Sang-Hee;Lee, Sun-Ah
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.17 no.1
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    • pp.7-11
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    • 2011
  • This research was conducted to compare the quality of paprika ($Capsicum$ $annuum$ L.) packed with PLA film and OPP/PE film pouches during storage at $6^{\circ}C$. Weight loss of paprika fruit was higher in OPP/PE film pouches, mold decay was shown in OPP/PE pouches after 6 days, but was not yet shown in PLA pouches. Hardness decreased in all packages. Soluble solid content was highest in PLA1 film pouch, vitamin C content was also highest in PLA2. Mold decay was not shown in PLA pouches after 9 day, but shown at calyx of paprika fruit in OPP/PE pouches. Off-flavor was strong in the OPP/PE pouches when the pouches were opened, decay was also serious in the same pouches after 15 days and thus quality was inadequate for sale in market.

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D609, an Inhibitor of Phosphatidylcholine-specific Phospholipase C, Inhibits Group IV Cytosolic Phospholipase A2

  • Kang, Mi Sun;Jung, Sung Yun;Jung, Kwang Mook;Kim, Seok Kyun;Ahn, Kyong Hoon;Kim, Dae Kyong
    • Molecules and Cells
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    • v.26 no.5
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    • pp.481-485
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    • 2008
  • As an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), D609 has been widely used to explain the role of PC-PLC in various signal transduction pathways. This study shows that D609 inhibits group IV cytosolic phospholipase $A_2$ ($cPLA_2$), but neither secretory $PLA_2$ nor a $Ca^{2+}$-dependent $PLA_2$. Dixon plot analysis shows a mixed pattern of noncompetitive and uncompetitive inhibition with $K_i=86.25{\mu}M$ for the $cPLA_2$ purified from bovine spleen. D609 also time- and dose-dependently reduces the release of arachidonic acid from a $Ca^{2+}$- ionophore A23187-stimulated MDCK cells. In the AA release experiment, $IC_{50}$ of D609 was ${\sim375}{\mu}M$, suggesting that this reagent may not enter the cells easily. The present study indicates that the inhibitory effects of D609 on various cellular responses may be partially attributable to the inhibition of $cPLA_2$.