• 제목/요약/키워드: ARTI

검색결과 67건 처리시간 0.033초

택사(Alismatis Rhizoma) trypsin inhibitor의 정제와 특성 (Purification and Characterization of Trypsin Inhibitor from Alismatis Rhizoma)

  • 박종옥;이인섭
    • 생명과학회지
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    • 제12권2호
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    • pp.151-157
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    • 2002
  • 한방재료의 하나인 택사(Alismatis Rhizoma, AR)로부터 단백성 trypsin inhibitor(TI)를 분리, 정제하여 특성을 조사하였다. 정제과정은 0-80.% 포화 황산암모늄을 이용한 염석법, DEAE-cellulose ion exchange chromatography, Sep-hadex G-150 chromatography 등을 이용하였다. 정제된 ARTI의 분자량을 gel filtration과 SDS-PAGE 한 결과 모두 약 23,000 Da으로 나타나 monomer로 되어 있는 것으로 나타났다. 온도안정성에 있어 0-6$0^{\circ}C$에서는 안정하였으나 그 이상의 온도에서는 약 35%가지 안정성이 떨어졌다. ARTI와 상품화된 soybean kunitz inhibitor의 저해능을 비교해 본 결과 ARTI 및 soybean inhibitor 각각의 농도가 0.071 $\mu$M, 1.7 $\mu$M일 때 0.025 g/$m\ell$ trypsin활성을 50% 정도 저해하는 것으로 나타났다. ARTI의 trypsin의 가수분해반응에 대한 저해형태는 비경쟁적 저해형인 것으로 나타났으며 km값은 0.81 $\mu$M이었다.

Kenalog modified by ionizing radiation induces intrinsic apoptosis mediated by elevated levels of reactive oxygen species in melanoma cancer

  • REMIGIUS AMBROSE KAWALA;FATUMA JUMAPILI RAMADHANI;HYO JIN CHOI;EUN-HEE LEE;CHUL-HONG PARK;BYUNG YEOUP CHUNG;HYOUNG-WOO BAI
    • Oncology Letters
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    • 제41권3호
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    • pp.1837-1850
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    • 2019
  • Kenalog is a synthetic glucocorticoid drug used to treat various cancers including ocular and choroidal melanoma. However, the drug achieves rarely sustainable results for patients. To overcome this difficulty, the structure of Kenalog was altered by ionizing radiation (IR) to develop a more effective anticancer agent for treatment of various skin cancers. The anticancer effect of modified Kenalog (Kenalog-IR) was assessed in melanoma cancer cells in vitro. The assessment of mitochondrial functions by MTT assay revealed significant inhibition of melanoma cancer cell viability by Kenalog-IR compared to Kenalog. Moreover, Kenalog-IR-induced apoptotic cell death was associated with the intrinsic mitochondrial pathway by triggering the release of intrinsic apoptosis molecules through activation of caspase-related molecules in concentration and time-dependent manners. Furthermore, it was observed that Kenalog-IR-induced apoptosis was associated with the generation of reactive oxygen species (ROS) with increased G2/M cell cycle arrest. Collectively, Kenalog-IR may be a potential suppressor of skin-related cancer in particular melanoma cancer.

Different Physiological Response to Salt in Salt Tolerant Rice Mutants Induced by Gamma-Mutagenesis

  • Jang, Duk-Soo;Song, Mira;Kim, Sun-hee;Kim, Jin-Baek;Kim, Sang Hoon;Ha, Bo-Keun;Kang, Si-Yong;Kim, Wook;Kim, Dong Sub
    • 방사선산업학회지
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    • 제5권3호
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    • pp.259-266
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    • 2011
  • When plants undergo stress, Reactive oxygen species (ROS) which remove bad elements such as mildew and virus is activated in plant body. However, if ROS is excessively increased, plant will be harmed itself by destruction of cell and signal system and phenomenon of lipid peroxidation. In order to identify content of lipid peroxidation and activity of some enzymes scavenging ROS, phenotypical and physiological analysis was performed with two mutant lines, Till-II-877 and Till-II-894, comparing with cv. Dongan (WT). In phenotype analysis, two mutant lines give to well-conditioned growth better than WT in since 5 days after salt treatment. In enzyme activities, there was a modest difference in the content of catalase (CAT) and peroxidase (POD) between Till-II-877 and Till-II-894, two mutant lines showed high levels in CAT contents than WT. However, they express low levels in POD contents. In MDA analysis, the content of Till-II-877 was higher than that of WT, but Till-II-894 was lower. This result indicates that two mutants have different mechanism against salt stress.

Differential Expression of C4H and F5H Genes in Rice (Oryza sativa L.) after Gamma-irradiation

  • Park, Young-Mi;Chae, Hyo-Seok;Chung, Byung-Yeoup;Kim, Jae-Sung;Kim, Jin-Hong;Wi, Seung-Gon;An, Byung-Chull;Cho, Jae-Young
    • 환경생물
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    • 제24권2호
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    • pp.155-159
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    • 2006
  • To reveal effects of gamma-irradiation with various doses on the expressions of C4H and F5H genes, the transcription levels of OsC4HL and OsF5HL were investigated in leaves and stems of two rice cultivars, Ilpoombyeo and IR-29, after the irradiation with 5, 10, 50, or 100 Gy for 4 h. In overall pattern of 24 h after the irradiation, the transcription levels of the two genes increased with the increasing doses of radiation in the leaves of both cultivars, except that of OsC4HL in IR-29. However, in the stems, the transcription level of OsF5HL increased in Ilpoombyeo and decreased in IR-29 dose-dependently, while that of OsC4HL decreased in Ilpoombyeo with the increasing doses of radiation and remained constant in IR-29. When the expressions of OsC4HL and OsF5HL were investigated in a time-course after the irradiation with 100 Gy, they reached their highest levels in the leaves of both cultivars 5 hand 72 h after the irradiation, respectively. Therefore, we suggest that the expressions of OsC4HL and OsF5HL, which involved in the same phenylpropanoid pathway, are differentially regulated during the post-irradiation period, showing different cultivar and tissue specificity. Furthermore, the dose dependency of the gene expressions is also discussed immediately after the irradiation.

택사(Alismatis Rhizoma)로부터 트립신 저해제의 정제와 특성 규명 및 이와 결합하는 단백질, 10-Formyltetrahydrofolate Dehydrogenase에 관한 연구 (Purification and Characterization of Trypsin Inhibitor from Alismatis Rhizoma and its Binding Protein, 10-Formyltetrahydrofolate Dehydrogenase)

  • 김지만;박종옥;신영희
    • 약학회지
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    • 제52권1호
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    • pp.79-84
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    • 2008
  • Alismatis Rhizoma has been used as diuretics and antiphlogistics in the Chinese oriental medicine. A trypsin inhibitor was isolated from Alismatis Rhizoma using DEAE ion exchange column, trypsin affinity column, and FPLC chromatography, and its activity and characteristics were studied. The purifed Alismatis Rhizoma trypsin inhibitor (ARTI) was estimated to be about 22 kDa. The sequence determination on N-terminal amino acid residues and 84 amino acid residues has been completed, yet no homology has been found with trypsin inhibitors reported at NCBI. ARTI did not show inhibitory activities on chymotrypsin and elastase, however it exhibited a significant inhibitory activity on bovine trypsin, and formed a complex with rat liver 10-formyltetrahydrofolate dehydrogenase.

cDNA cloning and expression pattern of Cinnamate-4-Hydroxylase in the Korean black raspberry

  • Baek, Myung-Hwa;Chung, Byung-Yeoup;Kim, Jin-Hong;Kim, Jae-Sung;Lee, Seung-Sik;An, Byung-Chull;Lee, In-Jung;Kim, Tae-Hoon
    • BMB Reports
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    • 제41권7호
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    • pp.529-536
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    • 2008
  • Cinnamate-4-hydroxylase (C4H) is a key enzyme in the phenylpropanoid pathway, which is responsible for synthesizing a variety of secondary metabolites that participate in development and adaptation. In this study, we isolated a full-length cDNA of the C4H gene from the Korean black raspberry (Rubus sp.) and found that this gene existed as a single gene. By comparing the deduced amino acid sequence of Rubus sp. C4H with other sequences reported previously we determined that this sequence was highly conserved among widely divergent plant species. In addition, quantitative real time PCR studies indicated that the C4H gene had a differential expression pattern during fruit development, where gene expression was first detected in green fruit and was then remarkably reduced in yellow fruit, followed by an increase in red and black fruit. To investigate the two peaks in expression observed during fruit development and ripening, we measured the flavonoid content. The content of the major flavanol of Korean black raspberry fruits was determined to be highest at the beginning of fruit development, followed by a gradually decrease according to the developmental stages. In contrast, the content of anthocyanins during the progress of ripening was dramatically increased. Our results suggest that the C4H gene in Korean black raspberry plays a role during color development at the late stages of fruit ripening, whereas the expression of C4H gene during the early stages may be related to the accumulation of flavanols.

국화 화색 돌연변이 품종 'ARTI-purple' 및 'ARTI-queen' 꽃잎 조직의 재분화와 신초형성에 미치는 식물생장호르몬의 영향 (The Effects of Plant Growth Regulators on Plant Regeneration and Direct Shoots Formation of Petal Explants of Chrysanthemum Flower Color Mutants Varieties, 'ARTI-purple' and 'ARTI-queen')

  • 이유미;강은정;성상엽;김상훈;하보근;김동섭;김진백;강시용
    • 원예과학기술지
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    • 제31권3호
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    • pp.359-365
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    • 2013
  • 국화는 전 세계적으로 가장 대중적인 화훼류 중의 하나로써, 최근 새로운 국화 품종들이 돌연변이육종을 통해 개발되고 있다. 그러나 현재까지 하나의 국화 원품종에서 유래한 돌연변이 품종들 간의 조직배양 조건 차이에 대한 보고는 전무하다. 따라서 본 연구에서는 2개의 국화 화색 돌연변이 품종 'ARTI-purple'과 'ARTI-queen'의 효율적인 재분화 조건을 비교하기 위해 수행되었다. 실험재료로 꽃봉오리 시기와 통상화 개화 이전 시기의 꽃잎을 이용하였고, 꽃의 조직에 따른 차이를 확인하기 위해 통상화 및 설상화를 구분하여 실험을 수행하였다. 국화 재분화를 위한 식물생장호르몬의 적정 조합을 찾기 위해 3% sucrose, 0.3% gelrite를 포함한 MS 배지에 BA, NAA, IAA 중 2가지 호르몬을 조합별로 첨가하여 실험을 수행하였다. 절편체는 일장 16시간, 온도 $25{\pm}1^{\circ}C$ 조건으로 배양하였으며, 재분화율 조사는 배양 후 4주 및 8주차에 실시하였다. 결과적으로, 가장 높은 재분화율은 두 품종 모두 최적 NAA와 BA의 호르몬 조합에서 꽃봉오리 시기의 통상화를 사용할 시 가장 높게 확인되었다. 재분화를 위한 식물생장호르몬의 최적 조합은 'ARTI-purple'의 경우 NAA $1.0mg{\cdot}L^{-1}$와 BA $0.5mg{\cdot}L^{-1}$로 47.9%의 재분화 효율을 보였으며, 'ARTI-queen'의 경우 NAA $2.0mg{\cdot}L^{-1}$와 BA $1.0mg{\cdot}L^{-1}$로 25.6%의 재분화 효율을 나타내었다. 재분화된 지상부는 1/2 MS배지에서 발근시켰으며, 기내 소식물체는 유리온실에서 성공적으로 순화되었다. 본 연구결과는 감마선을 사용한 돌연변이육종법에 의해 개발된 다양한 국화품종의 효율적인 재분화 시스템을 구축하는데 유용한 정보를 제공할 것이다.

Enhancement of the Chaperone Activity of Alkyl Hydroperoxide Reductase C from Pseudomonas aeruginosa PAO1 Resulting from a Point-Specific Mutation Confers Heat Tolerance in Escherichia coli

  • Lee, Jae Taek;Lee, Seung Sik;Mondal, Suvendu;Tripathi, Bhumi Nath;Kim, Siu;Lee, Keun Woo;Hong, Sung Hyun;Bai, Hyoung-Woo;Cho, Jae-Young;Chung, Byung Yeoup
    • Molecules and Cells
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    • 제39권8호
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    • pp.594-602
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    • 2016
  • Alkyl hydroperoxide reductase subunit C from Pseudomonas aeruginosa PAO1 (PaAhpC) is a member of the 2-Cys peroxiredoxin family. Here, we examined the peroxidase and molecular chaperone functions of PaAhpC using a site-directed mutagenesis approach by substitution of Ser and Thr residues with Cys at positions 78 and 105 located between two catalytic cysteines. Substitution of Ser with Cys at position 78 enhanced the chaperone activity of the mutant (S78C-PaAhpC) by approximately 9-fold compared with that of the wild-type protein (WT-PaAhpC). This increased activity may have been associated with the proportionate increase in the high-molecular-weight (HMW) fraction and enhanced hydrophobicity of S78C-PaAhpC. Homology modeling revealed that mutation of $Ser^{78}$ to $Cys^{78}$ resulted in a more compact decameric structure than that observed in WT-PaAhpC and decreased the atomic distance between the two neighboring sulfur atoms of $Cys^{78}$ in the dimer-dimer interface of S78C-PaAhpC, which could be responsible for the enhanced hydrophobic interaction at the dimer-dimer interface. Furthermore, complementation assays showed that S78C-PaAhpC exhibited greatly improved the heat tolerance, resulting in enhanced1 survival under thermal stress. Thus, addition of Cys at position 78 in PaAhpC modulated the functional shifting of this protein from a peroxidase to a chaperone.

In Vitro Plant Regeneration from Stolen Node Explant in Eremochloa Ophiuroides (Munro) Hack

  • Barampuram, Shyamkumar;Chung, Byung-Yeoup;Lee, Seung-Sik;An, Byung-Chull;Kim, Jae-Sung
    • Journal of Plant Biotechnology
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    • 제34권2호
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    • pp.161-166
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    • 2007
  • In vitro shoot regeneration and multiple shoot induction has been obtained from the stolen node explants in Eremochloa ophiuroides (Munro) Hack. The highest number of shoots ($10.66{\pm}0.21$) was observed from initial explants after one month culture duration on Murashige and Skoog (MS) medium containing 6-benzyladenine (BA: 0.5 mg/l). First generation shoot was excised and sub-cultured on the same fresh media for further multiplication of shoots. An enhanced number of second round shoots ($15.33{\pm}0.21$) was obtained compared to the initial culture media containing BA (0.5 mg/l). The number of shoots/stolon node was higher among all the concentrations of BA than kinetin (KN). In vitro regenerated shoots were successfully rooted in the phytohormone free MS medium. Plantlets generated with roots were transferred to pots containing compound mixture of soil and kept in green house conditions. Acclimatized plants showed 100% survival rate with normal morphology in green house conditions. The present study demonstrates the effect of explant and different plant growth regulators towards in vitro response in E. ophiuroides. Moreover, the study reveals the effect of cytokinin on induction of shoot number per stolen node explant in E. ophiuroides.