• Title/Summary/Keyword: 유도단백질

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Bee Venom Inhibits DU-145 Cell Proliferation Through Induction of Apoptosis (Bee Venom이 세포자멸사를 통해 DU-145 세포의 증식에 미치는 영향)

  • Hur, Keun-Young;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.28 no.3
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    • pp.111-119
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    • 2011
  • 목적 : 이 연구는 봉독이 세포자멸사 관련 단백질의 발현 조절을 통하여 세포자멸사를 유도하고 전립선 암세포주인 DU-145 세포의 성장을 억제하는지를 확인하고 해당 기전을 살펴보고자 하였다. 방법 : 봉독을 처리한 후 DU-145의 세포자멸사를 관찰하기 위해 TUNEL staining assay를 시행하였으며, 세포자멸사 조절단백질의 변동 관찰에는 western blot analysis를 시행하였다. 결과 : DU-145 세포에 봉독을 처리한 후, 세포자멸사의 유발, 세포자멸사 관련 단백질의 발현에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. DU-145 세포에서 봉독을 처리한 후 세포자멸사가 유도되어 세포성장이 억제되었다. 2. 세포자멸사 관련 단백질 중 분리된 pro-apoptotic proteins인 PARP, caspase-3, caspase-9은 유의한 증가를 나타내었다. 3. 세포자멸사 관련 단백질 중 분리된 anti-apoptotic proteins인 Bcl-2, p-AKT, XIAP, cIAP2는 유의한 감소를, MMP2, MMP13은 유의한 증가를 나타내었다. 결론 : 이상의 결과는 봉독이 인간 전립선 암세포주인 DU-145의 세포자멸사를 유발함으로써 전립선암세포 증식억제 효과가 있음을 입증한 것으로 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다.

Inhibitory Effects of Herb and Seeds of Oenothera odorata on the Melanogenesis (월견초 및 월견자의 멜라닌 생성 억제효과)

  • Choi, Mi-Eun;Woo, Won-Hong;Mun, Yeun-Ja
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.1
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    • pp.149-157
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    • 2010
  • 목적 : 월견초는 다량의 불포화 지방산인 리놀렌산과 감마 리놀렌산을 함유하고 있으며, 천식성 기침이나 아토피피부염에 효능이 있는 것으로 알려져 있다. 본 연구는 월견초의 전초와 종자 추출물의 피부 멜라닌 합성에 대한 억제 효과를 조사하였다. 방법 : B16F10 멜라닌세포주를 이용하여 멜라닌, tyrosinase 활성 및 세포생존율을 측정하였다. 또한 멜라닌 합성- 관련효소인 tyrosinase, TRP-1, TRP-2의 단백질발현과 $\alpha$-MSH를 처리하여 색소침착을 유도 한 뒤 단백질 발현을 조사하였다. 결과 : 월견자는 B16F10 세포의 멜라닌 합성을 $5\;{\mu}g/ml$$10\;{\mu}g/ml$ 농도에서 각각 대조군의 81.3%, 68.3%로 억제하였고 tyrosinase의 활성도 이와 유사하게 억제하였다. 멜라닌 합성-관련효소들의 단백질발현을 관찰한 결과 월견초와 월견자는 tyrosinase 발현을 억제하였으며 TRP-1과 TRP-2의 발현에는 영향을 주지 않았다. 특히 $\alpha$-MSH에 의한 과색소 유도 시 tyrosinase 발현이 현저하게 감소되었으며, 월견자의 멜라닌 합성 억제 효과가 월견초 보다 높게 나타났다. 결론 : 이상의 연구 결과 월견자는 멜라닌세포의 tyrosinase 단백질 발현과 tyrosinase 활성을 억제하여 멜라닌 생성을 감소시키는 것으로 사료된다.

유기용매 내성균주의 단백질 발현조사를 통한 heat shock와 oxidative stress의 유기용매내성과의 연관성

  • Choe, Seung-Tae;Lee, Ji-A;Bae, Gi-Jeong;Mun, Ja-Yeong;Jeong, Yeong-Gi;Ju, U-Hong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.253-256
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    • 2001
  • $3{\sim}5$ types of proteins were expressed by toluene and heat during $30{\sim}60$min. Generally it is reported that proteins below 10kDa function as transcription factor. In this study we certified that 7kDa was induced by organic solvent and the rate of expression was 2 folds at $30{\sim}45$min.

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High-level Expression of Human Procaspase-9 in Escherichia coli and Purification of its GST-tagged Recombinant Protein (대장균을 이용한 세포사멸 유도 단백질 caspase-9의 발현에 관한 연구)

  • Seong, Yeong-Mo;Han, Cheol;Choe, Ju-Yeon;Park, Hyo-Jin;Seong, Geun-Hye;Nam, Min-Gyeong;Kim, Sang-Su;Kim, In-Gyeong;Gang, Seong-Man;Im, Hyang-Suk
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.216-222
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    • 2003
  • Human caspase-9, an essential apoptosis initiator protease, was excessively degraded when expressed in Escherichia coli under the conventional induction condition. To optimize the conditions for induction and develop a rapid purification method for obtaining significant amounts of wild-type procaspase-9, we expressed procaspase-9 as GST fusion in E. coli. The addition of 0.01 mM IPTG as an inducer to the bacterial culture and decreasing the culture temperature to 25oC improved the production of procasapse-9 protein by circumventing proteolytic degradation in E. coli. The wild-type procaspae-9 was purified to approximately 70% purity with relatively high yields using the method developed in this study. In addition, we found that GST-caspase-9 is autocatalytically cleaved after aspartic acid 315, which is the same site for processing in mammalian cells, during expression in E. coli.

Red Ginseng Ethanol Extract Suppressed Ag I/II-induced Up-expression of Inflammatory Mediators in RAW 264.7 Macrophages (홍삼에탄올추출물의 염증유발인자에 대한 억제효과)

  • Choi, Kyung-Min;Hwang, Seung-Mi;Lim, Ji-Ye;Ko, Eun-Sil;Park, Jong-Hyuk;Moon, Jung-Hye;Lee, Min-Jung;Jang, Ji-Eun;Cha, Jeong-Dan
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.158-163
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    • 2015
  • In this study, we investigated the effects of 40% ethanol extract of Red Ginseng (RGE) on the productions of inflammatory proteins in Antigen I/II (Ag I/II)-N, a recombinant protein isolated from Streptococcus mutans -stimulated in RAW 264.7 cells. RGE inhibited the expression of Ag I/II-N-induced pro-inflammatory mediators, both mRNA and protein synthesis levels, without any cytotoxic effects. Moreover, RGE significantly inhibited Ag I/II-N induced NF-κB translocation into the nucleus by preventing the degradation of inhibitor κB-α. In conclusion, RGE down regulates the expression of pro-inflammatory genes involved in the synthesis of NO and iNOS in Ag I/II-N-stimulated RAW 264.7 cells by suppressing NF-κB activity.

Effect of Epigallocatechin Gallate on Apoptosis in MDA-MB-231 Human Breast Cancer Cells (Epigallocatechin Gallate가 인체 유방암 세포인 MDA-MB-231의 세포사멸에 미치는 영향)

  • Hong, Eun-Jung;Kim, Woo-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.9
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    • pp.1114-1119
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    • 2008
  • Among the numerous polyphenols isolated from green tea, epigallocatechin gallate (EGCG) is a predominate and is considered to be a major therapeutic agent. To elucidate the mechanical insights of anti-tumor effect, EGCG was applied to human breast cancer MDA-MB-231 cells. We investigated the effect of EGCG on protein and mRNA expression of proteins related to cell apoptosis in MDA-MB-231 human breast cancer cell lines. We also identified caspase-3 activity. We cultured MDA-MB-231 cells in the presence of 0, 5, 10, and $20\;{\mu}M$ of EGCG. Protein and mRNA expression of bcl-2 were decreased dose-dependently in cells treated with EGCG. However, protein and mRNA expression of bax were increased (p<0.05). Caspase-3 activities were increased dose-dependently in cells treated with EGCG. These results suggest that EGCG induces cell apoptosis by increase of caspase activity through decreasing of protein and mRNA expression of bcl-2 and increasing of protein and mRNA expression of bax.

Expression of a Human Histone H1.5 in Transgenic Tobacco Cultured Cells (담배 배양세포에서 인간 히스톤 단백질 H1.5의 발현)

  • Kim, Kee-Yeun;Kwon, Suk-Yoon;Song, Jae-Young;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.31 no.2
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    • pp.175-178
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    • 2004
  • Transgenic tobacco (Nicotiana tabacum L. cv Bright Yellow-2) cell lines expressing a human histone H1.5 (referred to as hH1.5), which suppress collagen-induced rheumatoid arthritis, were developed under the oxidative stress-inducible peroxidase (SWPA2) promoter. Tobacco BY-2 cells were transformed by Agrobacterium-mediated method. The kanamycin-resistant calli were selected on the modified MS medium containing 150mg/L kanamycin and 300mg/L claforan. Transgenic cell lines were confirmed by PCR and northern blot analysis. Recombinant hH1.5 (rhH1.5) protein (42 kDa) was also detected by Western blot analysis, showing a different molecular weight of human hH1.5 (32 kDa). These results suggested that a hH1.5 gene was properly introduced in tobacco cultured cells under the control of SWPA2 promoter. The further characterization of rhH1.5 protein remains to be studied.

Changes in Functional Groups of Protein by Lipid Deterioration in the Biological System of Rice Bran (미강 저장 중 지방의 산패에 따라 생성된 산화 지질이 단백질의 기능기에 미치는 영향에 관한 연구)

  • Song, Yeong-Ok;Choi, Hong-Sik
    • Korean Journal of Food Science and Technology
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    • v.22 no.5
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    • pp.596-601
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    • 1990
  • The effects of peroxidized lipid on the protein in the biological system of rice bran was studied by determining the changes in the content of functional groups under two different storage conditions. One stored at controlled atmosphere of $35^{\circ}C$ with relative humidity 65% and the other one was exposed to the air of $25^{\circ}-30^{\circ}C$ with relative humidity 70-90%. The lipid peroxidation started after the lipolysis was almost completed. The autoxidation occurred much faster in the bran exposed to the air than that stored in the controlled atmosphere. Substantial changes in the physiochemical characteristics were observed in all of the major functional groups in both of the samples. The content of sulfhydryl and available lysine decrease·1 as lipid peroxidation progressed. Protease activity was lost almost completely. Protein solubility and in vitro digestibility also decreased during storage. The lipid peroxidation and contents of major protein functional groups were significantly correlated (p<0.05) and the correlation coefficients were higher than -0.8, for the both of the sample. peroxidized lipid was found to deteriorate protein in the biological system as well.

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Relationship between RNA- and Protein-Synthesis and Cell Wall Acidification in Auxin-Mediated Elongation of Sunflower Hypocotyls (해바라기 하배축의 오옥신 유도 신장에서 RNA 및 단백질의 합성과 세포벽 산성화의 관계)

  • 조형택
    • Journal of Plant Biology
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    • v.35 no.2
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    • pp.107-116
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    • 1992
  • The roles of RNA- and protein-synthesis and $H^{+}$ excretion in 1AA ($10\;\mu\textrm{M}$)-induced elongation were investigated using abraded hypocotyl segments of sunflower (Helianthus annuus L.). The response of elongation initiated about 13 min after IAA treatment. Removal of cuticle, acting as diffusion barrier for inhibitors, by mechanical abrasion of hypocotyl segments enhanced the effect of inhibitors markedly, but the degree of abrasion for the saturated effect of inhibition was different among inhibitors. The elongation induced by 1M was completely inhibited when cycloheximide ($10\;\mu\textrm{M}$) was applied to abraded hypocotyl segments as shortly as 4 min before the onset of the growth response (= 10 min after administration of IAA). Cordycepin ($200\;\mu\textrm{M}$) prevented completely 1AA-induced elongation when applied as shortly as 19 min before the onset of the growth response (=5 min before administration of 1AA). Vanadate (1 mM) inhibited both lAA-induced elongation and medium acidification via lAA-induced $H^{+}$ excretion to apoplast. Cycloheximide and cordycepin also prevented lAA-induced $H^{+}$ excretion strongly. However, inhibition by cycloheximide of lAA-induced elongation was not alleviated by acidifying the cell wall to pH 4.5. The results indicate that, a few minutes before the initiation of growih, protein synthesis is demanded for the initiation of 1AA-induced elongation and the $H^{+}$ excretion to cell wall, and that the H+ excretion, even though it may be necessary for elongation, does not seem to bring about acid growth simply through acidifying cell wall.l wall.

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