• Title/Summary/Keyword: AP-$2{\alpha}$

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CK2 phosphorylates AP-2α and increases its transcriptional activity

  • Ren, Kaiqun;Xiang, Shuanglin;He, Fangli;Zhang, Wenfeng;Ding, Xiaofeng;Wu, Yanyang;Yang, Liping;Zhou, Jianlin;Gao, Xiang;Zhang, Jian
    • BMB Reports
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    • v.44 no.7
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    • pp.490-495
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    • 2011
  • Transcription factor AP-$2{\alpha}$ involves in the process of mammalian embryonic development and tumorigenesis. Many studies have shown that AP-$2{\alpha}$ functions in association with other interacting proteins. In a two-hybrid screening, the regulatory subunit ${\beta}$ of protein casein kinase 2 ($CK2{\beta}$) was identified as an interacting protein of AP-$2{\alpha}$; we confirmed this interaction using in-vitro GST pull-down and in-vivo co-immunoprecipitation assays; in an endogenous co-immunoprecipitation experiment, we further found the catalytic subunit ${\alpha}$ of protein casein kinase 2 ($CK2{\alpha}$) also exists in the complex. Phosphorylation analysis revealed that AP-$2{\alpha}$ was phosphorylated by CK2 kinase majorly at the site of Ser429, and such phosphorylation could be blocked by CK2 specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) in a dose-dependent manner. Luciferase assays demonstrated that both $CK2{\alpha}$ and $CK2{\beta}$ enhanced the transcription activity of AP-$2{\alpha}$; moreover, $CK2{\beta}$ increased the stability of AP-$2{\alpha}$. Our data suggest a novel cellular function of CK-2 as a transcriptional co-activator of AP-$2{\alpha}$.

Isolation and Chemical Analysis of Potent Anti-Complementary Polysaccharides from Fruiting Bodies of the Fomes fomentarius (말굽버섯 자실체에서 분리한 항보체 활성 다당체의 화학적 분석)

  • Park, Jung-Keun;Park, Kwe-Won;Shin, Kwang-Soon;Lee, Chang-Muk;Seok, Soon-Ja;Kim, Jeong-Bong;Koo, Bon-Sung;Han, Bum-Soo;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.198-206
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    • 2013
  • The five anti-complementary polysaccharides (MFKF-NP, MFKF-AP1${\alpha}$, ${\beta}$, and MFKF-AP2${\alpha}$, ${\beta}$) were separated from hot water extracts of fruiting bodies of Fomes fomentarius by two subsequent column chromatography using DEAE-sepharose FF and Concanavalin A-sepharose 4B. The order of anti-complementary activity was MFKF-AP1${\beta}$ > MFKF-AP1${\alpha}$ > MFKF-AP2${\alpha}$ > MFKF-AP2${\beta}$ > MFKF-NP > Polysaccharide Krestine (PSK). Especially, MFKF-AP1${\beta}$ among those showed the most excellent anti-complementary activity (70% of ITCH50 value at $20{\mu}g/ml$). The monosaccharide composition analysis by gas chromatography indicates that MFKF-AP1${\alpha}$ and ${\beta}$ are a kind of homoxylan consisted mainly of xylose above 97%. Molecular weight of MFKF-AP1${\beta}$, major anti-complementary polysaccharide, was estimated to be about 12,000 by high performance liquid chromatography (HPLC). After the incubation of the serum with MFKF-AP1${\beta}$ in the presence or absence of $Mg^{++}$ and $Ca^{++}$ ions, its anti-complementary activity was investigated. This result indicated that MFKF-AP1${\beta}$ seems to be activator both on the classical and the alternative pathway of complement activation.

Efficiency of Antioxidants on Types of Substrate Systems (기질 system의 유형에 따른 항산화제의 효과에 관한 연구)

  • 김찬희;안명수
    • Korean journal of food and cookery science
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    • v.14 no.5
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    • pp.560-565
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    • 1998
  • The purpose of this study was to investigate any differences in the efficiency of various antioxidants for the three types of substrates such as corn oil in water (O/W) emulsion, water in com oil (W/O) emulsion, and bulky corn oil. ${\alpha}$-Tocopherol (${\alpha}$-Toc) at 0.01 or 0.02%, ascorbic acid (AsA), ascorbyl palmitate (AP), and BHT at 0.02% were added separately to the prepared O/W emulsion, W/O emulsion, and bulk oil, and their antioxidative effects were compared. The mixture of ${\alpha}$-Toc ind AsA or AP at the level of 0.02% also was tested to observe any synergistic effect. Oxidation was made by storing at 42${\pm}$1$^{\circ}C$ for 25 days and the oxidative stability was determined by peroxide value and conjugated dienoic acid with time fluctuation of storage. The results were as follows: 1. In case of O/W emulsion, the order of antioxidative effect was AP> ${\alpha}$-Toc+AP>${\alpha}$-Toc+AsA>AsA>BHT. 2. In case of W/O emulsion, the order of antioxidative effect was AsA>AP>${\alpha}$-Toc+AsA>BHT. ${\alpha}$-Toc+AP mixture showed the prooxidant effect rather than synergistic effect. 3. In case of bulk oil, the order of antioxidative effect was AsA>AP>${\alpha}$-Toc+AsA>${\alpha}$-Toc+AP\ulcornerBHT. Therefore, AsA, a hydrophilic antioxidant, was more effective in W/O emulsion system than in O/W emulsion system, while the opposite trend was found in AP, a lipophilic antioxidant. AsA, a hydrophilic antioxidant, was more efficient in bulk oil of anhydrous substrate. ${\alpha}$-Toc showed prooxidant effects in all substrates.

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Anti-Inflammatory Effect of Fermented Artemisia princeps Pamp in Mice

  • Joh, Eun-Ha;Trinh, Hien-Trung;Han, Myung-Joo;Kim, Dong-Hyun
    • Biomolecules & Therapeutics
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    • v.18 no.3
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    • pp.308-315
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    • 2010
  • Essential oil-excluded Artemisia princeps Pamp var Ssajuarissuk (AP) was fermented with Lactobacillus brevis K-1, which was isolated from cabbage Kimchi, and the anti-inflammatory effects of AP and fermented AP (FAP) on lipopolysaccharide (LPS)-induced inflammatory response in peritoneal macrophages were investigated. AP and FAP inhibited LPS-induced TNF-$\alpha$, IL-$1{\beta}$, COX-2, iNOS and COX-2 expression, as well as NF-${\kappa}B$ activation. AP and FAP also reduced ear thickness, inflammatory cytokine (TNF-$\alpha$, IL-$1{\beta}$ and IL-6) expression and NF-${\kappa}B$ activation with 12-O-tetradecanoylphorbol-13-acetate (TPA) induced dermatitis in mice. Furthermore, AP and FAP also reduced exudate volume, cell number, protein amount, inflammatory cytokines (TNF-$\alpha$, IL-$1{\beta}$ and IL-6) expression and NF-${\kappa}B$ activation in carrageenan-induced air pouch inflammation in mice. The inhibitory effects of FAP were more potent than those of non-fermented AP. Based on these findings, we propose that FAP can improve inflammatory diseases, such as dermatitis, by inhibiting the NF-${\kappa}B$ pathway.

Serum concentrations of α2-macroglobulin, α1-antitrypsin, and C-reactive protein in dogs with suspected acute pancreatitis

  • Park, Soyoung;Kim, Hakhyun;Kang, Ji-Houn;Kang, Byeong-Teck;Yang, Mhan-Pyo
    • Korean Journal of Veterinary Research
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    • v.59 no.1
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    • pp.9-15
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    • 2019
  • In canine acute pancreatitis (AP), inappropriate release and activation of zymogen proteases within the pancreas results in the consumption of serum antiproteases. The aim of this study was to examine whether the serum concentrations of ${\alpha}_2$-macroglobulin (A2MG), ${\alpha}_1$-antitrypsin (A1AT), and C-reactive protein (CRP) differ between dogs with AP and healthy dogs. Twenty healthy dogs and 20 dogs with AP were included in this study. Concentrations of A2MG, A1AT, and CRP were measured in the sera of healthy dogs and dogs diagnosed with AP. Serum A2MG and A1AT concentrations were significantly lower in dogs with AP than in healthy dogs, whereas the serum CRP concentration was significantly higher. In addition, the concentrations of A2MG and A1AT were significantly higher in AP survivors than in AP non-survivors, while the CRP concentration was significantly lower. However, in both AP survivors and non-survivors, the CRP concentrations showed a negative correlation with A2MG concentrations but not with A1AT. These findings indicate that serum antiproteases and CRP concentrations might be associated with the mortality rate of AP in dogs.

Molecular cloning and characterization of novel human JNK2 (MAPK9) transcript variants that show different stimulation activities on AP-1

  • Wang, Pingzhang;Xiong, Ying;Ma, Chuan;Shi, Taiping;Ma, Dalong
    • BMB Reports
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    • v.43 no.11
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    • pp.738-743
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    • 2010
  • The c-Jun $NH_2$-terminal kinase (JNK) signaling pathway participates in many physiological functions. In the current study we reported the cloning and characterization of five novel JNK2 transcript variants, which were designated as $JNK2\alpha3$, $JNK2\alpha4$, $JNK2\beta3$, $JNK2\gamma1$ and $JNK2\gamma2$, respectively. Among them, $JNK2\alpha4$ and $JNK2\gamma2$ are potential non-coding RNA because they contain pre-mature stop codons. Both $JNK2\alpha3$ and $JNK2\beta3$ contain an intact kinase domain, and both encode a protein product of 46 kDa, the same as those of $JNK2\alpha1$ and $JNK2\beta1$. $JNK2\gamma1$ contains a disrupted kinase domain and it showed a disable function. When over-expressed in mammalian cells, $JNK2\alpha3$ showed higher activity on AP-1 than that of $JNK2\beta3$ and $JNK2\gamma1$. Furthermore, $JNK2\alpha3$ and $JNK2\beta3$ showed different levels of substrate phosphorylation, although they both could promote the proliferation of 293T cells. Our results further demonstrate that JNK2 isoforms preferentially target different substrates and may regulate the expression of various target genes.

Chemical Analysis of Acidic Proteo-heteroglycans with Anti-complementary Activity from the Hot-Water Extract of Fomitella fraxinea (장수버섯 자실체로부터 분리한 항보체 활성 단백다당체의 화학적 분석)

  • Yoon, Sang-Hong
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.502-510
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    • 1998
  • The hot-water extract of fruiting bodies in Fomitella fraxinea had potent anti-complementary activities. After fractionation of water-soluble polysaccharides by DEAE-Sephadex A-25 column chromatography, major anti-complementary activity was concentrated into the FF-AP1 among three polysaccharides (FF-NP, FF-AP1, FF-AP2). FF-AP1 was fractionated into $FF-AP1{\alpha}$ and $FF-AP1{\beta}$ obtained from the adsorbed fraction and unadsorbed fraction by affinity chromatography using a ConA-sepharose 4B column, respectively. $FF-AP1{\beta}$, which exihibited the highest anti-complementary activities had an IR absorption peak of $890cm^{-1}$, and a M.W. of about 15,000 (gel filtration). Anti-complementary activity of FF-AP1 decreased greatly by pronase treatment and periodate oxidation. $FF-AP1{\beta}$ responsible for potent anti-complemenary activities of Fomitella fraxinea was an acidic protein-containing heteroglycan consisted of 48% glucose, 13% mannose, and 12% galactose as major component sugars, 9.6% protein, 6% uronic acids.

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Cooperativity of ${\alpha}$- and ${\beta}$-Subunits of Group II Chaperonin from the Hyperthermophilic Archaeum Aeropyrum pernix K1

  • Kim, Jeong-Hwan;Lee, Jin-Woo;Shin, Eun-Jung;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.212-217
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    • 2011
  • ${\alpha}$ and ${\beta}$-subunits (ApCpnA and ApCpnB) are group II chaperonins from the hyperthermophilic archaeum Aeropyrum pernix K1, specialized in preventing the aggregation and inactivation of substrate proteins under conditions of transient heat stress. In the present study, the cooperativity of ${\alpha}$- and ${\beta}$-subunits from the A. pernix K1 was investigated. The ApCpnA and ApCpnB chaperonin genes were overexpressed in E. coli Rosetta and Codonplus (DE3), respectively. Each of the recombinant ${\alpha}$- and ${\beta}$-subunits was purified to 92% and 94% by using anionexchange chromatography. The cooperative activity between purified ${\alpha}$- and ${\beta}$-subunits was examined using citrate synthase (CS), alcohol dehydrogenase (ADH), and malate dehydrogenase (MDH) as substrate proteins. The addition of both ${\alpha}$- and ${\beta}$-subunits could effectively protect CS and ADH from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively, and MDH from thermal inactivation at $80^{\circ}C$C and $85^{\circ}C$. Moreover, in the presence of ATP, the protective effects of ${\alpha}$- and ${\beta}$-subunits on CS from thermal aggregation and inactivation, and ADH from thermal aggregation, were more enhanced, whereas cooperation between chaperonins and ATP in protection activity on ADH and MDH (at $85^{\circ}C$) from thermal inactivation was not observed. Specifically, the presence of both ${\alpha}$- and ${\beta}$- subunits could effectively protect MDH from thermal inactivation at $80^{\circ}C$ in an ATP-dependent manner.

Antiasthmic Effect of Fermented Artemisia princeps in Asthmic Mice Induced by Ovalbumin

  • Bae, Eun-Ah;Min, Sung-Won;Lee, Bo-Mi;Kim, Nam-Jae;Baek, Nam-In;Han, Eun-Joo;Chung, Hae-Gon;Kim, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1554-1557
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    • 2007
  • Artemisia princeps Pampanini (AP) was fermented with Bifidobacterium infantis K-525 and its antiasthmic effect investigated. AP and fennented AP (FAP) reduced the IgE level in the blood of ovalbumin-induced asthmic mice. Moreover, FAP reduced the IgE, proinflammatory cytokine IL-6, and IL-4 levels in the trachea, as well as in the lung of the experimental asthmic mice, whereas AP only reduced the IgE and IL-6 levels in the lungs. Nonetheless, AP and FAP both inhibited the mRNA expression of IL-6 and TNF-${\alpha}$ in IgE-induced RBL-2H3 cells. The in vivo antiasthmic effect of FAP was more potent than that of AP. Therefore, these findings suggest that the enhanced antiasthmic effect of AP after bifidus fermentation was possibly due to the regulation of the proinflammatory cytokine biosynthesis of IL-6 and TNF-${\alpha}$.

Protective Effects of Ethanol Extract from Aster Yomena on Acute Pancreatitis (쑥부쟁이 에탄올 추출물의 급성췌장염 보호 효과)

  • Seo, Sang Wan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.2
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    • pp.109-115
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    • 2019
  • Aster Yomena (AY) has been used in traditional medicine to treat diseases such as obesity, hyperlipidemia, atherosclerosis, diabetes and osteoarthritis. However, protective effect of AY on acute pancreatitis (AP) has not been reported. The present study examined the anti-inflammatory effects of an ethanol extract of AY on cerulein-induced AP. AP was induced in mice by intraperitoneally injecting cerulein ($50{\mu}g/kg$) hourly for 6 times. 70% ethanol extract of AY (0.1, 0.2, and 0.5 g/kg) was orally administered for 1 week before acute pancreatitis induction. The mouse was killed at 6 hours after the final cerulein injection. The pancreas and lung were rapidly removed for histological examination and myeloperoxidase (MPO) assay. Blood samples were taken to determine serum amylase and lipase activity. In addition real-time reverse transcription-polymerase chain reaction (RT-PCR) was also performed to investigate mRNA expression of proinflammatory cytokines such as $TNF-{\alpha}$. $IL-1{\beta}$, and IL-6. Administration of AY significantly ameliorated pancreatic weight to body weight ratio, histological damages and MPO activity during AP. In addition, AY inhibited the serum amylase and lipase activity during AP. Also, mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 were inhibited by AY against AP. Our results revealed that pre-treatment of AY reduces the severity of cerulein-induced AP. Therefore, AY may have a protective effect drug against AP.