• Title/Summary/Keyword: catalytic factor

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Analysis of Gene Expression Modulated by Indole-3-carbinol in Dimethylbenz[a]anthracene-induced Rat Mammary Carcinogenesis

  • Kang, Jin-Seok;Park, Han-Jin;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.5 no.3
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    • pp.222-229
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    • 2009
  • Our previous finding that pre-initiation treatment of indole-3-carbinol (I3C) represents a chemopreventive effect in dimethylbenz[a]anthracene (DMBA)-induced mammary carcinogenesis has prompted us to test the global expression of genes at an early stage. Rats were continuously fed 300 ppm I3C in their diet at 6 weeks of age and were injected with DMBA at 7 weeks of age, and were sacrificed at 8 weeks of age. Global gene expression analysis using oligonucleotide microarrays was conducted to detect altered genes in DMBA- or DMBA plus I3C-treated mammary glands. Altered genes were identified by fold changes of 1.2 and by t-test (P<0.05) from the log ratios of the hybridization intensity of samples between control (Group 1) and DMBA (Group 2), and from those of samples between DMBA (Group 2) and DMBA plus I3C (Group 3). From these genes, we chose altered genes that were up- or down-regulated by DMBA treatment and recovered to the control level by I3C treatment. For early stage of carcinogenesis, I3C treatment induced the recovery to normal levels of several genes including cell cycle pathway (cyclin B2, cell division cycle 2 homolog A), MAP signaling pathway (fibroblast growth factor receptor 1, platelet derived growth factor receptor, beta polypeptide), and insulin signaling (protein phosphatase 1, regulatory (inhibitor) subunit 3B and flotillin 2), which were up-regulated by DMBA treatment. In addition, I3C treatment induced the recovery to normal levels of several genes including those of MAPK signaling (transforming growth factor, beta receptor 1 and protein phosphatase 3, catalytic subunit, beta isoform), which were down-regulated by DMBA treatment. These results suggest that the targeting of these genes presents a possible approach for chemoprevention in DMBA-induced mammary carcinogenesis.

Identification of Calcium/Calmodulin-Dependent Phosphatase as the Dephosphorylating Enzyme of IgE-Dependent Histamine-Releasing Factor in RBL-2H3 (RBL-2H3 세포에서 IgE-depnedent Histamine-releasing Factor의 탈인산화 효소에 관한 연구)

  • Hwang Sun-Ok;Lee Kyunglim
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.189-193
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    • 2005
  • IgE-dependent histamine-releasing factor(HRF) was initially described as a secretagogue for secretion of histamine from IgE+ basophils from a subset of allergic donors. Previously, we identified that S98 residue of HRF was phosphorylated using anti-HRFpS98 antibody which specifically recognizes the phosphorylated serine residue of HRF and HRFS98A mutant construct. In vitro kinase assay, only wild type HRF was phosphorylated by PKC, and S98A HRF was not affected by PKC. In this study, we attempted to characterize the phosphatase which specifically dephosphorylates HRF by immunoprecipitation and pull-down assay. In RBL-2H3 cells, HRF interacted only with calcineurin (also called as PP2B, calcium/calmodulin-dependent phosphatase) but not with PP1 or PP2A. The results suggest that HRF is most likely dephosphory-lated by calcineurin.

Contribution of a Low-Barrier Hydrogen Bond to Catalysis Is Not Significant in Ketosteroid Isomerase

  • Jang, Do Soo;Choi, Gildon;Cha, Hyung Jin;Shin, Sejeong;Hong, Bee Hak;Lee, Hyeong Ju;Lee, Hee Cheon;Choi, Kwan Yong
    • Molecules and Cells
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    • v.38 no.5
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    • pp.409-415
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    • 2015
  • Low-barrier hydrogen bonds (LBHBs) have been proposed to have important influences on the enormous reaction rate increases achieved by many enzymes. ${\Delta}^5$-3-ketosteroi isomerase (KSI) catalyzes the allylic isomerization of ${\Delta}^5$-3-ketosteroid to its conjugated ${\Delta}^4$-isomers at a rate that approache the diffusion limit. Tyr14, a catalytic residue of KSI, has been hypothesized to form an LBHB with the oxyanion of a dienolate steroid intermediate generated during the catalysis. The unusual chemical shift of a proton at 16.8 ppm in the nuclear magnetic resonance spectrum has been attributed to an LBHB between Tyr14 $O{\eta}$ and C3-O of equilenin an intermediate analogue, in the active site of D38N KSI. This shift in the spectrum was not observed in Y30F/Y55F/D38N and Y30F/Y55F/Y115F/D38N mutant KSIs when each mutant was complexed with equilenin, suggesting that Tyr14 could not form LBHB with the intermediate analogue in these mutant KSIs. The crystal structure of Y30F/Y55F/Y115F/D38N-equilenin complex revealed that the distance between Tyr14 $O{\eta}$ and C3-O of the bound steroi was within a direct hydrogen bond. The conversion of LBHB to an ordinary hydrogen bond in the mutant KSI reduced the binding affinity for the steroid inhibitors by a factor of 8.1-11. In addition, the absence of LBHB reduced the catalytic activity by only a factor of 1.7-2. These results suggest that the amount of stabilization energy of the reaction intermediate provided by LBHB is small compared with that provided by an ordinary hydrogen bond in KSI.

Occurrence of Leaf Spot Caused by Stemphylium lycopersici on Cirsium setidens in Korea (Stemphylium lycopersici에 의한 고려엉겅퀴 점무늬병의 발생)

  • Choi, Hyo-Won;Kim, Seok Gu;Hong, Sung Kee;Lee, Young Kee;Lee, Jae Guem;Kim, Hyo Won;Lee, Eun Hyeong
    • The Korean Journal of Mycology
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    • v.44 no.3
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    • pp.201-205
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    • 2016
  • In August 2015, leaf spot symptoms were observed on Korean gondre thistle (Cirsium setidens) in Youngwol, Korea. During the early stage, the symptoms appeared as one or more small gray-brown to brown spots on plant leaves. The spots showed extensive enlargement over time and eventually became large dark brown to black lesions on the whole leaf. Stemphylium species were consistently isolated from affected leaves. All isolates were identified as S. lycopersici, S. solani, or S. xanthosomatis based on morphological and cultural characteristics. The isolates were confirmed as S. lycopersici based on a multilocus sequence analysis using the ribosomal internal transcribed spacer (ITS) region, elongation factor 1, GAPDH (glyceraldehyde-3-phosphate dehydrogenase), and the noncoding region between the vacuolar membrane ATPase catalytic subunit A gene and a gene involved in vacuolar biogenesis. Pathogenicity was tested by spore suspension inoculation on wounded or unwounded gondre leaves. The lesions were observed on inoculated leaves within 3 days after inoculation, regardless of wound. To our knowledge, this is the first report of the leaf spot on gondre thistle caused by S. lycopersici in Korea or elsewhere.

Effects of dietary Antrodia cinnamomea fermented product supplementation on metabolism pathways of antioxidant, inflammatory, and lipid metabolism pathways-a potential crosstalk

  • Lee, M.T.;Lin, W.C.;Lin, L.J.;Wang, S.Y.;Chang, S.C.;Lee, T.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.7
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    • pp.1167-1179
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    • 2020
  • Objective: This study was conducted to fathom the underlying mechanisms of nutrition intervention and redox sensitive transcription factors regulated by Antrodia cinnamomea fermented product (FAC) dietary supplementation in broiler chickens. Methods: Four hundreds d-old broilers (41±0.5 g/bird) assigned to 5 groups were examined after consuming control diet, or control diet replaced with 5% wheat bran (WB), 10% WB, 5% FAC, and 10% FAC. Liver mRNA expression of antioxidant, inflammatory and lipid metabolism pathways were analyzed. Prostaglandin E2 (PGE2) concentration in each group were tested in the chicken peripheral blood mononuclear cells (cPBMCs) of 35-d old broilers to represent the stress level of the chickens. Furthermore, these cells were stimulated with 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) and lipopolysaccharide (LPS) to evaluate the cell stress tolerance by measuring cell viability and oxidative species. Results: Heme oxygenase-1, glutathione S-transferase, glutamate-cysteine ligase, catalytic subunit, and superoxide dismutase, and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) that regulates the above antioxidant genes were all up-regulated significantly in FAC groups. Reactive oxygen species modulator protein 1 and NADPH oxygenase 1 were both rather down-regulated in 10% FAC group as comparison with two WB groups. Despite expressing higher level than control group, birds receiving diet containing FAC had significantly lower expression level in nuclear factor-kappa B (NF-κB) and other genes (inducible nitric oxide synthase, tumor necrosis factor-α, interleukin-1β, nucleotide-binding domain, leucine-richcontaining family, pyrin domain-containing-3, and cyclooxygenase 2) involving in inflammatory pathways. Additionally, except for 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase that showed relatively higher in both groups, the WB, lipoprotein lipase, Acetyl-CoA carboxylase, fatty acid synthase, fatty acid binding protein, fatty acid desaturase 2 and peroxisome proliferator-activated receptor alpha genes were expressed at higher levels in 10% FAC group. In support of above results, promoted Nrf2 and inhibited NF-κB nuclear translocation in chicken liver were found in FAC containing groups. H2O2 and NO levels induced by LPS and AAPH in cPBMCs were compromised in FAC containing diet. In 35-d-old birds, PGE2 production in cPBMCs was also suppressed by the FAC diet. Conclusion: FAC may promote Nrf2 antioxidant pathway and positively regulate lipid metabolism, both are potential inhibitor of NF-κB inflammatory pathway.

Hydrogen Peroxide Sensitive Biosensors Based on Mugwort-Peroxidase Entrapped in Carbon Pastes (탄소반죽에 쑥 과산화효소를 고정한 과산화수소 감응 바이오센서)

  • Yoon, Kil Joong
    • Applied Chemistry for Engineering
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    • v.26 no.5
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    • pp.624-629
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    • 2015
  • A biosensor including the homogenized tissue of mugwort embedded in carbon paste, which senses hydrogen peroxide, was constructed and its electrochemical properties were validated using voltammetry. The good linearity of Hanes-Woolf plot implied that the reduction reaction of substrate was catalyzed by mugwort peroxidase at the electrode surface. Also the small value of symmetry factor, 0.28, indicated that electrochemical kinetics of the sensor is very sensitive to the change of electrode potential. Many experimental results collected above proved that the dissociation of hydrogen peroxide is dependent on the catalytic power of mugwort peroxidase qualitatively and quantitatively at the surface of the mugwort electrode. It is our firm belief that the marketed HRP can be replaced with mugwort tissue.

Isolation of Grb2-Shc Domain Binding Inhibition Component from Agastache rugosa (배초향으로부터 Grb2-Shc domain 결합저해 물질의 분리)

  • Lee, Eun-Sook;Ahn, Byung-Tae;Lee, Sae-Bom;Kim, Hyae-Kyeong;Bok, Song-Hae;Jeong, Tae-Sook
    • Korean Journal of Pharmacognosy
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    • v.30 no.4
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    • pp.404-408
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    • 1999
  • SH2 domains and their associated catalytic or noncatalytic proteins constitute critical signal transduction targets for drug discovery. Grb2 associates with phosphotyrosine sites of the activated receptors or Shc via their SH2 domain to link receptor tyrosine kinases to ras signalling. Blocking of the Grb2-Shc complex may be to intervene the oncogenic signal transduction pathways and to develop a new antitumor drug. In the search for blockers of Grb2 SH2-Shc interaction, Lutein, a family of carotenoids, was isolated from the extract of the leaf of Agastache rugosa O. Kuntze as SH2 domain antagonists. The $IC_{50}$ of Lutein against Grb2-Shc binding was $6.8\;{\mu}M$.

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Sequence Analysis of $\beta$-Xylosidase Gene from Bacillus stearothermophilus (Bacillus stearothermophilus $\beta$-Xylosidase 유전자의 염기 서열 결정 및 분석)

  • 오현주;최용진
    • Microbiology and Biotechnology Letters
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    • v.22 no.2
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    • pp.134-142
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    • 1994
  • The neucleotide sequences of the xylA gene encoding $\beta $-xylosidase of Bacillus stearothermophilus and is its flanking regions were datermined. Three open reading frame(ORFs) were found, one of which(ORF1) appeared to code for the $\beta $-xylosidase. The 1830 base pair ORF1 encoded 609 amino acids starting from a TTG initiation codon. The molecular weight deduced from the nucleotide sequence(68 KD) was in agreement with that estimated by SDS-polyacrylamide gel electrophoresis of the purified enzyme(66 KD). The Shine-Dalgarno sequence(5'-AGGAGG-3') was found 11 bp upstream of the initiation codon. Further 15 bp upstream, there observed a potential transcription initiation signals. The putative -10 sequence(CATAAT) and -35 sequence(TTGTTA) coresponded closely to the consensus sequences for Bacillus subtilis RNA polymerase with major sigma factor. The guanine-plus-cytosine content of the coding region of the xylA gene was 56mol% while that of the third position of the codons was 63 mol%. Based on the comparison with the amino acid sequences of several other carbohydrate degrading enzymes, two conserved regions, possibly participating in the catalytic mechamism of $\beta $-xylosidase xylA, were identified in 278-298 and 329-350 regions of the translated xylA gene. The nucleotide sequence of the xylA was found to exhibit no homology to any other genes so far reproted.

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Characteristics of LPG Fuel Reforming using Plasma Technology (플라즈마를 이용한 LPG연료 개질 특성연구)

  • Kim, Changup
    • Transactions of the Korean hydrogen and new energy society
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    • v.26 no.1
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    • pp.1-7
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    • 2015
  • In this study, characteristics of reforming process of automotive liquefied petroleum gas (LPG) fuel using plasma reactor are investigated. Because plasma reformer technology has advantages of a fast start-up and wide fuel/oxidizer ratio of operation, and reactor size is smaller and more simple compared to typical combustor and catalytic reactor, plasma reforming is suitable to the on-board vehicle reformer. To evaluate the characteristics of the reforming process, parametric effect of $O_2/C$ ratios, reactant flow rate and metal form on the process were investigated. In the test of varying $O_2/C$ ratio from partial oxidation to stoichiometry combustion, conversion of LPG was increased but selectivity of $H_2$ decreased. The optimum condition of $O_2/C$ ratio for the highest $H_2$ yield was determined to be around 1.0 for 20~50 lpm, and 1.35 for 100 lpm. Specific energy density (SED) was major factor in reforming process and higher SED leads to higher $H_2$ yield. And metal form in the reformer increased $H_2$ yield of about 34 % as compared to the case of no metal form. The result can be a guide to map optimal condition of reforming process.

Regulation of Apoptosis by Nitrosative Stress

  • Kim, Ki-Mo;Kim, Peter K.M.;Kwon, Young-Guen;Bai, Se-Kyung;Nam, Woo-Dong;Kim, Young-Myeong
    • BMB Reports
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    • v.35 no.1
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    • pp.127-133
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    • 2002
  • Nitrosative stress can prevent or induce apoptosis. It occurs via S-nitrosylation by the interaction of nitric oxide (NO) with the biological thiols of proteins. Cellular redox potential and non-heme iron content determine S-nitrosylation. Apoptotic cell death is inhibited by S-nitrosylation of the redox-sensitive thiol in the catalytic site of caspase family proteases, which play an essential role in the apoptotic signal cascade. Nitrosative stress can also promote apoptosis by the activation of mitochondrial apoptotic pathways, such as the release of cytochrome c, an apoptosis-inducing factor, and endonuclease G from mitochondria, as well as the suppression of NF-${\kappa}B$ activity. In this article we reviewed the mechanisms whereby S-nitrosylation and nitrosative stress regulate the apoptotic signal cascade.