• 제목/요약/키워드: bPDI

검색결과 31건 처리시간 0.021초

소포체 스트레스에 대한 Protein Disulfide Isomerase의 세포보호효과 (Bombyx mori Protein Disulfide Isomerase (bPDI) Protects Sf9 Cells from Endoplasmic Reticulum (ER) Stress)

  • 구태원;윤은영;김성완;최광호;강석우;권기상;권오유
    • 생명과학회지
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    • 제17권8호통권88호
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    • pp.1129-1134
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    • 2007
  • bPDI가 ER내 misfolding 단백질의 생성을 제한함으로써 곤충변역과 관계하는지를 해석하기 위하여 bPDI가 과발현(overexpression)되는 곤충세포주와 이와 반대로 bPDI가 억제발현(knock-down)되는 곤충세포주를 제작하여 bPDI가 곤충면역에 관련하는지를 해석하였다. bPDI가 과발현되는 세포주 (Sf9-bPDI)는 정상세포주(Sf9)나 pIZT/V5-His 벡터만 도입된 세포주(Sf9-pIZT)에 비하여 생존율이 30% 이상 높았지만, bPDI의 전사체 발현이 억제된 세포주(Sf9-bPDI-dsRNA)는 오히려 정상세포주나 pIZT/V5-His 벡터만 도입된 세포주에 비하여 생존율이 약 15%낮았다. 이와 같은 결과로써, bPDI는 ER내 misfolding 단백질의 생성을 제한함으로써 곤충의 ERSE과 밀접하게 관련할 것이라 추정할 수 있었다.

Baculovirus Expression and Biochemical Characterization of the Bombyx mori Protein Disulfide Isomerase (bPDI)

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Park, Kwang-Ho;Hwang, Jae-Sam;Kwon, O-Yu;Kang, Seok-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • 제7권2호
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    • pp.127-131
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    • 2003
  • Protein disulfide isomerase (PDI) found in the endoplasmic reticulum (ER) catalyzes disulfide bond exchange and assists in protein folding of newly synthesized proteins. PDI also functions as a molecular chaperone and has been found to be associated with proteins in the ER. In addition, PDI functions as a subunit of two more complex enzyme systems: the prolyl-4-hydroxylase and the triacylglycerol transfer proteins. A cDNA that encodes protein disulfide isomerase was previously isolated from Bombyx mori (bPDI), in which open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and an ER retention signal of the KDEL motif at its C-terminal, and we report its functional characterization here. This putative bPDI cDNA is expressed in insect Sf9 cells as a recombinant proteins using baculovirus expression vector system. The bPDI recombinant proteins are successfully recognized by antirat PDI antibody, and shown to be biologically active in vitro by mediating the oxidative refolding of reduced and scrambled RNase. This suggests that bPDI may play an important role in protein folding mechanism of insects.

Enzymatic activity of Endoplasmic Reticulum Oxidoreductin 1 from Bombyx mori

  • Park, Kwanho;Yun, Eun-Young;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • 제37권1호
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    • pp.15-20
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    • 2018
  • Most proteins produced in the endoplasmic reticulum (ER) of eukaryotic cells fold via disulfide formation (oxidative folding). Oxidative folding is catalyzed by protein disulfide isomerase (PDI) and PDI-related ER protein thiol disulfide oxidoreductases (ER oxidoreductases). In yeast and mammals, ER oxidoreductin-1s (ERO1s) supply oxidizing equivalent to the active centers of PDI. We previously identified and characterized the ERO1 of Bombyx mori (bERO1) as a thioredoxin-like protein that shares primary sequence homology with other ERO1s. Here we compare the reactivation of inactivated rRNase and sRNase by bERO1, and show that bERO1 and bPDI cooperatively refold denatured RNase A. This is the first result suggesting that bERO1 plays an essential role in ER quality control through the combined activities of bERO1 and bPDI as a catalyst of protein folding in the ER and sustaining cellular redox homeostasis.

Expression of Recombinant Human Stem Cell Factor (hSCF) Protein using Bombyx mori Protein Disulfide Isomerase (bPDI)

  • Kim, Sung-Wan;Yun, Eun-Young;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Lee, Kwang-Gill;Kwon, O-Yu;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • 제21권2호
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    • pp.151-155
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    • 2010
  • Protein disulfide isomerase (PDI) catalyzes the oxidation of disulfides and the isomerizatiob of incorrect disulfides in new polypeptides during folding in the oxidizing environment of the endoplasmic reticulum (ER). To increase recombinant protein hSCF (human stem cell factor) production, we have developed expression system using the Bombyx mori PDI (bPDI) as a fusion partner. bPDI gene fusion was found to improve the production of recombinant hSCFs. Thus, we conclude that bPDI gene fusion will be very useful for the large-scale production of biologically active recombinant proteins.

누에 배양세포로부터 분리한 Protein Disulfide Isomerase 유전자의 발현 특성 (Molecular Characterization of a Bombyx mori Protein Disulfide Isomerase(bPDI))

  • 구태원;윤은영;황재삼;강석우;권오유
    • 생명과학회지
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    • 제11권5호
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    • pp.415-422
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    • 2001
  • Many secreted proteins have disulfide bonds that are important for their structure and function. Protein disulfide isomerase (PDI, EC 5.3.1.4.), an enzyme that catalyzes the formation and rearrangement of thiol/disulfide exchange reactions, is a resident of the endoplasmic reticulum (ER). The subcellular localization and its function as catalyst of disulfide bond formation in the biosynthesis of secretory and cell membrane proteins suggest that PDI plays a key role in the secretory pathway. We have isolated a cDNA encoding protein disulfide isomerase from Bombyx mori(bPDI). It has been characterized under ER stress conditions (dominantly induced by calcium ionophore A23187, tunicamycin and DTT), which is known to cause an accumulation of unfolded proteins in the ER. Furthermore, It has also been examined for tissue distribution(pronounced at the fat body), hormonal regulation (juvenile hormone, insulin and juvenile +transferrin; however, it is not effected by transferrin alone), and the effect of exogenous bacteria (peak at 16 h after infection) on the bPDI mRNA expression. The results suggest that bPDI is a member of the ER stress protein group, and it may play an important role in exogenous bacterial infection in fat body, and that homones regulate its expression.

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소나무 수피 알칼리성 아황산염-안트라퀴논 증해폐액이 사료결착제로서 펠렛 내구성과 가금의 생산성에 미치는 영향 (Effects of pine Bark Spent Liquor Prepared by Alkaline Sulfite-Anthraquinone Cooking as a Pellet Binder on Pellet Durability and Performance of Broiler Chicks or Laying Hens)

  • K. S. Ryu;H. L. Li;S. P. Mun;H. J. Song
    • 한국가금학회지
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    • 제28권2호
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    • pp.91-98
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    • 2001
  • 소나무 수피로부터 alkaline sulfite-anthraquinone cooking을 통하여 생산된 Pine Bark Spent Liquor(PBSL)의 펠렛 내구성(ellet durability index(PDI)과 사료결착재로서 가능성을 구명하기 위하여 육계와 산란계 사료에 첨가·급여하였다. 실험1에서는 실용계 사료인 대조구, 소맥 10%와 (W10), 20% 첨가구(W20), PBSL 0.25, 0.50, 1.00, 2.00, 3.00% 첨가구, 두 종류 시판용 사료결착제 0.1, 0.2%(CPB A와 CPB B), W10+0.1% CPB A or B로 14개 처리구를 4반복으로 하여 펠렛 내구성 조사를 시행하였다. PDI는 PDI tester(Oriental motors, Japan)로 측정하였다. 실험2에서는 1일령 육계수컷 200수를 무첨가구인 대조구외에 실험 1에서 최적으로 판단되는 처리구인 W10, PBSL 0.5%, 0.2% CPB A and B의 5개 처리구를 4반복으로 배치하여 시행하였다. 기초사료는 사육전기에 3,169와 3,149kca./kg ME와 21% CP를 급여하였고, 사육후기에는 3,192와 3,172 kcal/kg ME와 19% CP사료를 급여하였다. 증체량, 사료섭취량, 사료요구율은 주간별로 5주간 측정하였으며 실험 종료시에 장내 혐기적인 미생물 수를 조사하였다. 실험 3에서 24주령 로만 갈색계 450수를 이용하여 처리구는 실험 2와 동일하게 8주간 시행하였다. 처리구당 반복은 18수씩 5반복으로 90수 전체 450수를 이용하였다. 사료는 대상에너지 2,720 kcal/kg와 CP는 18.5%로 하였다. 처리구당 산란율, 난중, 사료섭취량, 사료요구율, 계란의 품질을 주간 별로 조사하였으며, 실험종료시에 혈액의 성상을 조사하였다. 실험 1에서 PBSL 0.5%, W10 0.5%, CPB A, B 0.2% 처리구의 PDI는 95.9, 95.9, 95.8. and 95.7로서 대조구에 비하여 현저하게 높았다.(P<0.05). 그러므로 이러한 결과는 실험2와 3에 적용하였다. 실험 2에서 PBSL 처리구의 증체량은 대조두와 통계적인 차이는 없었지만 W10과 CPB A와는 현저하게 차이가 있었다. (P<0.05). 사료요구율은 PBSL이나 다른 사료결착제 처리구에서 대조구에 비하여 개선되는 경향을 보였다.

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Effect of a Bombyx mori Protein Disulfide Isomerase on Production of Recombinant Antibacterial Peptides

  • Goo, Tae-Won;Kim, Seong-Wan;Choi, Kwang-Ho;Kim, Seong-Ryul;Kang, Seok-Woo;Park, Seung-Won;Yun, Eun-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • 제26권2호
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    • pp.119-123
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    • 2013
  • The insect baculovirus expression vector system (BEVS) is useful for producing biologically active recombinant proteins. However, the overexpression of heterologous proteins using this system often results in misfolded proteins and the formation of protein aggregates. To overcome this limitation, we developed a versatile baculovirus expression and secretion system using Bombyx mori protein disulfide isomerase (bPDI) as a fusion partner. bPDI gene fusion was found to improve the secretions and antibacterial activities of recombinant nuecin and enbocin proteins. Thus, we conclude that bPDI gene fusion is a useful addition to BEVS for the large-scale production of bioactive recombinant proteins.

Bombyx mori protein disulfide isomerase enhances the production of nuecin, an antibacterial protein

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Choi, Kwang-Ho;Kang, Seok-Woo;Kwon, Ki-Sang;Yu, Kweon;Kwon, O-Yu
    • BMB Reports
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    • 제41권5호
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    • pp.400-403
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    • 2008
  • The insect baculovirus expression vector system (BEVS) is useful for producing biologically active recombinant proteins. However, the overexpressions of foreign proteins using this system often results in misfolded proteins and the formation of protein aggregates. To overcome this limitation, we developed a versatile baculovirus expression and secretion system using Bombyx mori protein disulfide isomerase (bPDI) as a fusion partner. bPDI gene fusion was found to improve the secretions and antibacterial activities of recombinant nuecin proteins. Thus, we conclude that bPDI gene fusion is a useful addition to BEVS for the large-scale production of bioactive recombinant proteins.

교통사고로 유발된 경항통 환자의 근에너지기법(MET)의 효과에 대한 임상적 연구 (The Clinical Studies for Muscle Energy Techniques on Patients with Nuchal Pain Caused by Traffic Accidents)

  • 전태동;이한길;홍서영;허동석;윤일지
    • 한방재활의학과학회지
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    • 제19권1호
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    • pp.125-134
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    • 2009
  • Objectives : The purpose of this study is to find out the effects of muscle energy techniques on nuchal pain caused by traffic accidents. Methods : The 20 patients were divided into 2 groups: group A was treated except muscle energy techniques and group B was treated with muscle energy techniques. Both groups were treated with acupunture treatment, physical theraphy and herbal medication. Patients were evaluated by McGill Pain Questionnaire-Short Form(SF-MPQ), Pain Disability Index(PDI) and Neck Disability Index(NDI) in traffic accident patients. Results : 1. Both Group were significantly decreased in sensory of SF-MPQ, VAS, PDI and NDI after 7 days of treatment. 2. Group B compared with the Group A was significantly decresed in VAS, PDI after 7 days of treatment. Conclusions : We found out that muscle energy techniques is considered to be effective and useful on nuchal pain caused by traffic accidents.

Structure Analysis of the Full Length PDI Genomic DNA Isolated from Bombyx mori

  • Kim, Sung-Wan;Goo, Tae-Won;Yun, Eun-Young;Park, Kwang-Ho;Hwang, Jae-Sam;Kang, Seok-Woo;Kwon, O-Yu
    • 한국잠사학회:학술대회논문집
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    • 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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    • pp.44-44
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    • 2003
  • Protein disulfide isomerase (PDI) is not only an isomerase catalyzing the formation of native disulfide bond(s) of nascent peptide, but also a molecular chaperone assisting chain folding. We have already reported the structure of a cDNA (bPDl) encoding PDI from Bombyx mori and the function of PDI as foldase in assisting protein folding. (omitted)

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