• 대한바이러스학회지
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• 제27권1호
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• pp.9-17
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• 1997

Hepatitis C virus (HCV) E2 protein is known to be one of putative envelope proteins. To develop a sensitive detection method for HCV infected tissues and cells, monoclonal antibodys (MAbs) to the E2 protein of HCV were prepared from mice immunized with recombinant baculovirus-expressing E2 protein (Bac-E2). Several hybridoma clones secreting various levels of MAb were isolated and isotypes of these MAb were determined. One clone (L.2.3.3) was used for ascites production and the E2-MAb was purified and characterized. The L.2.3.3 reacted well with both Bac-E2 and E. coli expressed glutathione-S-transferase-E2 (GST-E2) fusion proteins. Using HCV patient sera, E2 envelope protein was found to be localized in the cell membrane boundary both in CHO cells and insect cells which express HCV E2 protein. Similar result was obtained when same cells were treated with the MAb L.2.3.3. These results demonstrated that Bac-E2 protein is capable of eliciting high titer antibody production in mice.

• 한국수산과학회지
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• 제50권6호
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• pp.684-693
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• 2017

We investigated methods for extracting histidine-containing low-molecular-weight (LMW) peptides such as anserine, carnosine and histidine from the edible meat of tuna byproducts. Extracts were treated by several methods including heat treatment ($80^{\circ}C$, 10 min), DOWEX ion exchange (IEC), ultrafiltration (UF), and carboxymethyl (CM)-cellulose column chromatography (IEC+CMC); then the levels of protein, total iron, histidine, carnosine, and anserine were measured. Extracts treated with IEC+CMC using CM-cellulose were analyzed for total iron, protein, histidine, and anserine content, which were $6.27{\pm}0.26mg/mL$, $5.20{\pm}0.21{\mu}g/mL$, 0.80 mg/mL, 0.208 mg/mL, and 4.40 mg/mL, respectively, in yellowfin tuna; and $9.05{\pm}0.82mg/mL$, $4.06{\pm}0.20{\mu}g/mL$, 1.62 mg/mL, 0.012 mg/mL, and 7.28 mg/mL in bigeye tuna. By comparison in IEC-UF treated extracts, protein, total iron, and histidine content decreased by 43%, 73%, and 27% in yellowfin and 0.4%, 54%, and 23% in bigeye tuna, wheres carnosine and anserine content increased by 22% and 17%, respectively. Freeze-dried (FD) extracts exhibited similar trends as non-dried extracts, i.e., dipeptide content increased with purification steps, whereas pro-oxidant (total iron and protein) content decreased. IEC+CMC treated FD extracts had the highest anserine, content, and the greatest reductuion in pro-oxidants.

• 미생물학회지
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• 제29권2호
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• pp.75-79
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• 1991

The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.

• 생명과학회지
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• 제32권2호
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• pp.86-93
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• 2022

Lupeol은 pentacyclic triterpene의 일종으로 다양한 질병에 약리 효과가 있는 것으로 보고되어 있으나, lupeol이 포도당 흡수에 미치는 영향은 아직 보고된 바 없다. 본 연구에서 3T3-L1 지방세포에서 포도당 흡수에 대한 lupeol의 효과를 조사하였다. 그 결과, Lupeol은 3T3-L1 지방세포에서 GLUT4를 원형질막으로 이동시켜 포도당 흡수를 촉진하였으며, 이는 PI3K/AKT 및 AMPK 경로의 활성화와 관련되어 있었다. PI3K/AKT 경로에서 lupeol은 PI3K를 활성화시키는 insulin receptor substrate 1의 인산화와 AKT의 인산화를 촉진하지만 비정형 단백질 키나아제 C isoforms ζ 및 λ의 인산화는 촉진하지 않았다. 반면, lupeol은 5 'AMP-activated protein kinase의 인산화를 촉진하였고, Lupeol의 의한 AMPK의 활성화는 원형질막-GLUT4의 발현과 세포내 포도당 흡수를 증가시키는 것으로 확인되었다. 3T3-L1 지방세포에서 lupeol에 의한 포도당 흡수 효과는 PI3K 억제제인 wortmannin 및 AMPK 억제제인 Compound C에 의해 억제됨을 통해 확인하였다. 본 연구 결과는 lupeol이 3T3-L1 지방세포에서 PI3K/AKT 및 AMPK 경로를 통해 원형질막 GLUT4의 발현을 자극함으로써 인슐린 감수성을 증가시켜 포도당 흡수를 촉진할 수 있음을 제시하고 있다.

• 대한화장품학회지
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• 제41권3호
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• pp.263-268
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• 2015

본 연구는 오디씨 에탄올 추출물(MSE)의 멜라닌 합성 저해 효과를 밝히는 것이다. 먼저 MSE의 melan-a 세포를 이용한 멜라닌 합성 저해 실험결과, 독성을 보이지 않는 $10{\mu}g/mL$의 농도까지 멜라닌 합성 저해 효과를 나타내었다. 또한 tyrosinase, tyrosinase-related protein-1 (TRP-1) 단백질의 발현이 저해되었으며, extracelluar signal-regulated protein kinase (ERK)의 발현을 농도 의존적으로 증가시키는 MSE의 기전을 확인하였다. 뿐만 아니라, 제브라피쉬를 이용한 in vivo 모델의 실험에서도 색소 발생이 저해됨을 관찰하였다. 따라서 오디씨로 부터 획득한 에탄올 추출물이 ERK 단백질의 발현으로 인해 멜라닌 생합성을 억제할 수 있음을 밝혔다.

• 대한의생명과학회지
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• 제11권2호
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• pp.103-108
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• 2005

2-Deoxyribonolactone (dL) arises as a major DNA damage induced by a variety of agents, involving free radical attack and oxidation of C1'-deoxyribose in DNA. We investigated whether dL lesions can be repaired in mammalian cells and the mechanisms underlying the role of DNA polymerase $\beta$ in processing of dL lesions. Pol $\beta$ appeared to be trapped by dL residues, resulting in stable DNA-protein cross-links. However, repair DNA synthesis at site-specific dL sites occurred effectively in cell-free extracts, but predominantly accompanied by long-patch base excision repair (BER) pathway. Reconstitution of long-patch BER demonstrated that FEN1 was capable of removing the displaced flap DNA containing a 5'-dL residue. Cellular repair of dL lesions was largely dependent on the DNA polymerase activity of Pol $\beta$. Our observations reveal repair mechanisms of dL and define how mammalian cells prevent cytotoxic effects of oxidative DNA lesions that may threaten the genetic integrity of DNA.

• 한국잡초학회지
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• 제10권3호
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• pp.227-232
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• 1990

The effects of alachlor [2-chloro-2', 6' diethyl-N-(methoxymethyl) acetanilide] treatment on nucleic acid, amino acid and protein synthesis were studied. The amide herbicide alachlor blocks the biosynthesis of the amino acids isoleucine, valine and aromatic amino acid in oat root tips. Nucleic acid was inhibited, but was not proportional to reduction in protein synthesis. $1{\times}10^{-4}M$ of alachlor treatment of oat roots inhibited 36% DNA synthesis, but DNA synthesis was not inhibited at $1{\times}10^{-5}M$. RNA synthesis was inhibited by $1{\times}10^{-5}M$ and $1{\times}10^{-4}M$ of alachlor 16 and 27%, respectively, while inhibition of protein synthesis did occur at same concentrations. Inhibition of protein synthesis also did not occur at concentration below $1{\times}10^{-4}M$ alachlor. It suggest that inhibition of protein sythesis caused significantly by alachlor($1{\times}l0^{-3}M$) result from secondary action.

• 한국식품영양과학회지
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• 제31권1호
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• pp.1-6
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• 2002

강낭콩을 가열한 후 B. subtilis ATCC 51189로 발효시켜 새로운 활성 단백질의 생성여부를 확인하였다. 생강낭콩의 수용성 단백질은 열처리에 의하여 감소하였으며, 발효과정에서 단백질의 양은 변화하지 않았으나, 새로운 아미노산이 합성되었다. 생강낭콩의 수용성 단백질(raw protein, RP)은 분자량 118 kDa의 PHA(phytohemagglutinin)의 전형적인 모습을 보였으나, 열처리한 단백질(heated protein, HP)의 경우, RP의 band는 사라지고, 저분자 peptide로 변화되었으며, 발효과정에 의하여 분자량 18.0 kDa의 새로운 단백질(fermented protein, FP)이 생성된 것을 확인하였다. 또한, RP는 128 HU, HP는 4HU, FP는 32HU의 적혈구응집효과를 보였으며, 위암세포(SNU-1)에 대하여 RP 및 FP는 비교적 높은 농도($IC_{50}$/ $\mu\textrm{g}$/mL)에서 항암활성을 보였다. 그리고, RP 및 FP의 경우 1 $\mu\textrm{g}$/mL에서 림프구 증식효과를 보였고, IFN-${\gamma}$, IL-12의 분비를 촉진하였다. 그러나 HP는 항암효과, 림프구 증식효과, cytokine의 분비촉진효과를 보이지 않았다. 따라서, 강낭콩 lectin은 가열에 의하여 활성물질이 파괴되나, B. subtilis에 의한 발효과정에서 새로운 cytokine 분비촉진물질이 생성되는 것을 확인할 수 있었다.

• Nutrition Research and Practice
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• 제9권6호
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• pp.606-612
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• 2015

BACKGROUND/OBJECTIVES: Several medicinal properties of Smilax china L. have been studied including antioxidant, anti-inflammatory, and anti-cancer effects. However, the antiobesity activity and mechanism by which the water-soluble fraction of this plant mediates its effects are not clear. In the present study, we investigated the lipolytic actions of the water-soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) in 3T3-L1 adipocytes. MATERIALS/METHODS: The wsSCLE was identified by measuring the total polyphenol and flavonoid content. The wsSCLE was evaluated for its effects on cell viability, lipid accumulation, glycerol, and cyclic adenosine monophosphate (cAMP) contents. In addition, western blot analysis was used to evaluate the effects on protein kinase A (PKA), PKA substrates (PKAs), and hormone-sensitive lipase (HSL). For the lipid accumulation assay, 3T3-L1 adipocytes were treated with different doses of wsSCLE for 9 days starting 2 days post-confluence. In other cell experiments, mature 3T3-L1 adipocytes were treated for 24 h with wsSCLE. RESULTS: Results showed that treatment with wsSCLE at 0.05, 0.1, and 0.25 mg/mL had no effect on cell morphology and viability. Without evidence of toxicity, wsSCLE treatment decreased lipid accumulation compared with the untreated adipocyte controls as shown by the lower absorbance of Oil Red O stain. The wsSCLE significantly induced glycerol release and cAMP production in mature 3T3-L1 cells. Furthermore, protein levels of phosphorylated PKA, PKAs, and HSL significantly increased following wsSCLE treatment. CONCLUSION: These results demonstrate that the potential antiobesity activity of wsSCLE is at least in part due to the stimulation of cAMP-PKA-HSL signaling. In addition, the wsSCLE-stimulated lipolysis induced by the signaling is mediated via activation of the ${\beta}$-adrenergic receptor.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3817-3825
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• 2015

Background: The human protein methyl-transferase DOT1L catalyzes the methylation of histone H3 on lysine 79 (H3K79) at homeobox genes and is also involved in a number of significant processes ranging from gene expression to DNA-damage response and cell cycle progression. Inhibition of DOT1L activity by shRNA or small-molecule inhibitors has been established to prevent proliferation of various MLL-rearranged leukemia cells in vitro, establishing DOT1L an attractive therapeutic target for mixed lineage leukemia (MLL). Most of the drugs currently in use for the MLL treatment are reported to have low efficacy, hence this study focused on various natural compounds which exhibit minimal toxic effects and high efficacy for the target receptor. Materials and Methods: Structures of human protein methyl-transferase DOT1L and natural compound databases were downloaded from various sources. Virtual screening, molecular docking, dynamics simulation and drug likeness studies were performed for those natural compounds to evaluate and analyze their anti-cancer activity. Results: The top five screened compounds possessing good binding affinity were identified as potential high affinity inhibitors against DOT1L's active site. The top ranking molecule amongst the screened ligands had a Glide g-score of -10.940 kcal/mol and Glide e-model score of -86.011 with 5 hydrogen bonds and 12 hydrophobic contacts. This ligand's behaviour also showed consistency during the simulation of protein-ligand complex for 20000 ps, which is indicative of its stability in the receptor pocket. Conclusions: The ligand obtained out of this screening study can be considered as a potential inhibitor for DOT1L and further can be treated as a lead for the drug designing pipeline.