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유기인계 및 카바메이트계 농약의 고감수성 아세틸콜린에스테라이즈의 대량생산
김영미,조문제,김소미 한국응용생명화학회 2003 Applied Biological Chemistry (Appl Biol Chem) Vol.46 No.4
본 실험에서는 acetylcholinesterase(AChE, EC 3.1.1.7)를 이용한 간이 잔류농약 검사법에 필요한, 유기인계 및 카바메이트계 살충제에 대한 감수성이 증가된 AChE(MAChE)를 baculovirus를 이용하여 대량으로 생산하는 시스템을 구축하고 생산된 효소의 특성을 관찰하였다.한라산에서 채취한 초파리에서 AChE의 cDNA를 합성한 수 PCR을 이용하여 AChE의 lipid anchor 부분을 제거하고 site directed mutagenesis에 의해 E107Y, F368L, L408F의 염기서열을 변화시켜 재조합된 MAChE cDNA를 합성하였고 baculovirus vector에 삽입하여 대량생산을 시도하였다. 대량 증식에 필요한 조건으로 감염횟수가 네 번일 때, 그리고 세포수가 2×10^(6) cell/㎖일 때 세포의 증식과 효소의 활성이 극대화됨을 알 수 있었다. His tag을 붙여 Ni-NTA affinity column을 이용하여 MAChE를 정제하였으며, 정제된 효소는 실험조건하에서는 pH(3-10)와 온도(20-50℃)의 변화에 영향을 받지 않았다. 농약 추출액으로 methanol을 사용했을 때가 ethanol을 사용할 때 보다 효과적임을 알 수 있었다. 대표적인 유기인계와 카바메이트계 농약에 대한 저해율을 조사한 결과 재조합된 MAChE는 대만의 집파리 및 변형되지 않은 AChE에 비하여 전반적으로 농약에 대한 감수성이 높은 것으로 나타났다. For the simple rapid bioassay of organophosphorus and carbamate pesticide residues, a mass-production system of acetycholinesterase (AChE, EC 3.1.1.7, MAChE) using baculovirus and insect cell culture was constructed. The cDNA for AChE was synthesized from Drosophila melanogaster in Halla Mountain, the lipid anchor tail was removed by PCR and was used for the site-directed mutagenesis of three amino acid residues (E107Y, F368L, L408F). The mutated cDNA was inserted into the baculovirus vector and expressed in insect cells. Maximum cell growth and enzyme activity were reached when the cells (2×10^(6) cell/㎖) were infected four times at four-day-intervals. His-tag containing MAChE was prufied using Ni-NTA column and used for characterization. The activity was maintained under various pHs (3-10) and temperatures (20-50℃) under experimental conditions. As an extractionn solution for pesticides, methanol is more effective than ethanol. Against major organophosphate and carbamate pesticides, the MAChE showed better sensitivity than AChE and AChE from housefly (Taiwan).
Binding of Galectin-1 to Neutrophils Enhanced by Activation
Cho, Somi K.,Cho, Moonjae 한국응용생명화학회 2000 Journal of Applied Biological Chemistry (J. Appl. Vol.43 No.3
Human polymorphonuclear neutrophils undergo diaphoresis after a selectin-mediated rolling on the endothelial cells of the blood vessel wall. Extravasation is believed to be an integrin-mediated process. Galectin-1 is a small dimeric beta-galactoside-binding protein synthesized by the endotherial cells and present in the perivascular connective tissue. In this study we suggest the possible role of galectin-1 in extravasation of the activated neutrophils. MAL lectin binding study showed, that f-MetLeuPhe-activated neutrophils decrease surface sialylation and increase galectin-1 binding via exposure of new galectin-1 binding sites. Desialylated HL-60 cells also show the same decrease in MAL binding and increase in galectin-1 binding, an increase which was not observed in the presence of lactose. Galectin-1 blotting analysis detected two possible major ligands (approximately 120 and 160 kDa) of galectin-1 from the desialylated HL-60 cell lysates.
Binding of Galectin-1 to Neutrophils Enahnced by Activation
(Somi K . Cho,(Moon Jae Cho 한국응용생명화학회 2000 Journal of Applied Biological Chemistry (J. Appl. Vol.43 No.3
Human polymorphonuclear neutrophils undergo diaphoresis after a selectin-mediated rolling on the endothelial cells of the blood vessel wall. Extravasation is believed to be an integrin-mediated process. Galectin-1 is a small dimeric beta-galactoside-bindin
Kim, Chulwon,Cho, Somi K,Kim, Ki-Dong,Nam, Dongwoo,Chung, Won-Seok,Jang, Hyeung-Jin,Lee, Seok-Geun,Shim, Bum Sang,Sethi, Gautam,Ahn, Kwang Seok Rapid Science Publishers ; Kluwer Academic Publish 2014 Apoptosis Vol.19 No.4
<P>We have recently reported that β-caryophyllene oxide (CPO) can induce apoptosis, suppress tumor growth, and inhibit metastasis through the suppression of signal transducer and activator of transcription 3, PI3K/AKT/mTOR/S6K1 signaling cascades and ROS-mediated MAPKs activation. In the present study, we found that CPO potentiated the apoptosis induced by tumor necrosis factor α (TNFα) and chemotherapeutic agents, suppressed TNFα-induced tumor cell invasion, all of which are known to require NF-κB activation. We found that TNFα stimulated the expression of gene products involved in anti-apoptosis (IAP1, IAP2, Bcl-2, Bcl-xL, and survivin), proliferation (COX-2, cyclin D1, and c-Myc), invasion (MMP 9 and ICAM-1), and angiogenesis (VEGF) and that CPO treatment suppressed their expression. Because these gene products are also regulated by proinflammatory transcription factor NF-κB, we postulated that CPO may mediate its effects by modulating the NF-κB pathway. We found that CPO blocked both inducible and constitutive NF-κB activation in a wide variety of tumor cells. CPO was also found to inhibit the TNFα-induced degradation of IκBα through the inhibition of activation of IκBα kinase and p65 nuclear translocation and phosphorylation. Interestingly, CPO failed to potentiate the apoptotic effect induced by TNFα in p65 (-/-) cells as compared to the wild-type. Thus, overall, our results indicate that the inhibition of NF-κB is one of major mechanisms by which CPO enhances TNFα-induced apoptosis and suppresses invasion.</P>
Lim, Hee-Kyoung,Cho, Somi-K.,Park, Sang-Gyu,Cho, Moon-Jae The Korean Society for Applied Biological Chemistr 2010 Applied Biological Chemistry (Appl Biol Chem) Vol.53 No.3
Artemisia capillaris is a perennial herb that belongs to the family Campositae spp, and has been utilized in Korea for the treatment of abdominal pain, hepatitis, chronic liver disease, and coughing. The anti-inflammatory activity of the ethanol extract of A. capillaris (EtEAC) was investigated in this study. EtEAC did not affect the viability of various cells in a concentration range of $0-200\;{\mu}g/mL$. The DPPH radical scavenging ability of $10\;{\mu}g/mL$ of EtEAC was comparable to that of $10\;{\mu}g/mL$ of catechin. EtEAC exerted an anti-inflammatory effect on the mRNA expression levels of typical inflammatory cytokines (IL-6, $IL1{\beta}$) and cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Additionally, EtEAC reduced nitric oxide (NO) production via the downregulation of inducible nitric oxide synthase transcription, rather than by scavenging NO. EtEAC exerted its anti-inflammatory effects by suppressing the activation of p38 and ERK at $200\;{\mu}g/mL$ in LPS-stimulated RAW 264.7 cells.
( Hee Kyoung Lim ),( Somi K. Cho ),( Sang Gyu Park ),( Moon Jae Cho ) 한국응용생명화학회 2010 Applied Biological Chemistry (Appl Biol Chem) Vol.53 No.3
Artemisia capillaris is a perennial herb that belongs to the family Campositae spp, and has been utilized in Korea for the treatment of abdominal pain, hepatitis, chronic liver disease, and coughing. The anti-inflammatory activity of the ethanol extract of A. capillaris (EtEAC) was investigated in this study. EtEAC did not affect the viability of various cells in a concentration range of 0-200 μg/mL. The DPPH radical scavenging ability of 10 μg/mL of EtEAC was comparable to that of 10 μg/mL of catechin. EtEAC exerted an anti-inflammatory effect on the mRNA expression levels of typical inflammatory cytokines (IL-6, IL1β) and cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Additionally, EtEAC reduced nitric oxide (NO) production via the downregulation of inducible nitric oxide synthase transcription, rather than by scavenging NO. EtEAC exerted its anti-inflammatory effects by suppressing the activation of p38 and ERK at 200 μg/mL in LPS-stimulated RAW 264.7 cells.
KIM, So-Hyun,CHO, Somi K.,HYUN, Sun-Hee,PARK, Hae-Eun,KIM, Young-Suk,CHOI, Hyung-Kyoon Japan Society for Bioscience, Biotechnology, and A 2011 Bioscience, Biotechnology, and Biochemistry Vol.75 No.6
<P>Guava leaves were classified and the free radical scavenging activity (FRSA) evaluated according to different harvest times by using the <SUP>1</SUP>H-NMR-based metabolomic technique. A principal component analysis (PCA) of <SUP>1</SUP>H-NMR data from the guava leaves provided clear clusters according to the harvesting time. A partial least squares (PLS) analysis indicated a correlation between the metabolic profile and FRSA. FRSA levels of the guava leaves harvested during May and August were high, and those leaves contained higher amounts of 3-hydroxybutyric acid, acetic acid, glutamic acid, asparagine, citric acid, malonic acid, <I>trans</I>-aconitic acid, ascorbic acid, maleic acid, <I>cis</I>-aconitic acid, epicatechin, protocatechuic acid, and xanthine than the leaves harvested during October and December. Epicatechin and protocatechuic acid among those compounds seem to have enhanced FRSA of the guava leaf samples harvested in May and August. A PLS regression model was established to predict guava leaf FRSA at different harvesting times by using a <SUP>1</SUP>H-NMR data set. The predictability of the PLS model was then tested by internal and external validation. The results of this study indicate that <SUP>1</SUP>H-NMR-based metabolomic data could usefully characterize guava leaves according to their time of harvesting.</P>
Hee Kyoung Lim,Somi K. Cho,Sang Gyu Park,Moon Jae Cho 한국응용생명화학회 2010 Journal of Applied Biological Chemistry (J. Appl. Vol.53 No.3
Artemisia capillaris is a perennial herb that belongs to the family Campositae spp, and has been utilized in Korea for the treatment of abdominal pain, hepatitis, chronic liver disease, and coughing. The anti-inflammatory activity of the ethanol extract of
Kim, So-Hyun,K. Cho, Somi,Min, Tae-Sun,Kim, Yujin,Yang, Seung-Ok,Kim, Hee-Su,Hyun, Sun-Hee,Kim, Hana,Kim, Young-Suk,Choi, Hyung-Kyoon the Society for Free Radical Research Japan 2011 Journal of clinical biochemistry and nutrition Vol.48 No.3
<P>The ameliorating effects of Mango (<I>Mangifera indica</I> L.) flesh and peel samples on plasma ethanol level were investigated using a mouse model. Mango fruit samples remarkably decreased mouse plasma ethanol levels and increased the activities of alcohol dehydrogenase and acetaldehyde dehydrogenase. The <SUP>1</SUP>H-NMR-based metabolomic technique was employed to investigate the differences in metabolic profiles of mango fruits, and mouse plasma samples fed with mango fruit samples. The partial least squares-discriminate analysis of <SUP>1</SUP>H-NMR spectral data of mouse plasma demonstrated that there were clear separations among plasma samples from mice fed with buffer, mango flesh and peel. A loading plot demonstrated that metabolites from mango fruit, such as fructose and aspartate, might stimulate alcohol degradation enzymes. This study suggests that mango flesh and peel could be used as resources for functional foods intended to decrease plasma ethanol level after ethanol uptake.</P>
Song, Yeon Woo,Park, Sanggyu,Cho, Somi K.,Koh, Dongsoo,Cho, Moonjae The Korean Society for Applied Biological Chemistr 2013 Applied Biological Chemistry (Appl Biol Chem) Vol.56 No.6
Several naturally occurring or synthesized forms of chalcone have been shown to possess multiple biological properties, including antitumor activities. A novel synthetic flavonoid, the benzochalcone derivative DK-78, was administered with the anticancer drug doxorubicin to two breast cancer cell lines (MCF7-VN and MDA-MB-231), and was evaluated for a synergistic cytotoxic effect. DK-78 reduced the expression of mesenchymal marker proteins and reduced cell migration and attachment. Sequential treatment with DK-78 and doxorubicin showed synergistic effects.