• Title/Summary/Keyword: bPDI

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

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Choi, Kwang-Ho;Kang, Seok-Woo;Kwon, Ki-Sang;Kwon, O-Yu
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1129-1134
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    • 2007
  • In the previous our study, a cDNA that encodes protein disulfide isomerase from Bombyx mori (bPDI)was isolated and characterized. bPDI has an open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and ER (endoplasmic reticulum) retention signal of the KDEL motif at its C-terminal. Recent studies have demonstrated that misfolded proteins are accumulated in many diseases including Alzheimer’s, goiter, emphysema, and prion infections. bPDI was over-expressed or knock-downed in Sf9 cells to study the relationship between bPDI expression and protections against protein misfolding. bPDI gene was cloned in insect expression vector pIZT/V5-His for over-expression and bPDI double-stranded RNA (dsRNA) was generated for knock-down. Over-expression of bPDI significantly improved survival rate, but bPDI dsRNA transfection significantly reduced survival rate after 48 hours exposure. In mock-transfected or wild-type cells had no significant effect. The results support the view that bPDI is one of the important intracellular components for cell protect mechanism, especially, against ER stress such as protein misfolding.

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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    • v.7 no.2
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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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    • v.37 no.1
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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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    • v.21 no.2
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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.

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

  • 구태원;윤은영;황재삼;강석우;권오유
    • Journal of Life Science
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    • v.11 no.5
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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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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 (소나무 수피 알칼리성 아황산염-안트라퀴논 증해폐액이 사료결착제로서 펠렛 내구성과 가금의 생산성에 미치는 영향)

  • K. S. Ryu;H. L. Li;S. P. Mun;H. J. Song
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.91-98
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    • 2001
  • Three experiments were conducted to investigate the Pine Bark Spent Liquor (PBSL) inclusion, prepared by alkaline sulfite-anthraquinone cooking, on Pellet durability index (PDI) of practical diets and performance of broiler chicks and laying hens. Fourteen treatments with four replications were assigned for PDI test in Experiment 1. Control, 10% Wheat(W10), 20% Wheat(W20), 0.25, 0.50, 1.00, 2.00 and 3.00% PBSL, 0.1, 0.2% commercial pellet binder A(CPB A) or B(CPB B), W10 plus 0.1% CPB A or B were used for PDI test. PDI was measured by PDI tester (Oriental Motors, Japan). The control diet was based on corn and soybean meal with no wheat or pellet binders inclusion. The PDI of the PBSL or other commercial pellet binder treatments were significantly higher than control groups(P<0.05). It was shown 95.9, 95.9, 95.8, and 95.7 in W10, 0.5% PBSL, 0.2% CPB A or B treatments, respectively. Thus, those treatments were applied to Experiments 2 and 3. In experiment 2, 200 male broiler chicks (Cobb$\times$Cobb) were allocated to the control, W10, PBSL 0.5%, 0.2% CPB A and B with four replications. Starter diets contained 3,169, 3,149 kcal/kg ME and 21% CP, and finisher diets were fed at the level of 3,192, 3,172 kcal/kg ME and 19W% CP. Weight gain, feed intake, feed conversion ratio (FCR) were weekly measured for 5 wk and the number of intestinal anaerobes were examined at the end of experiment. The weight gain of chicks fed PBSL was not significantly greater than control groups, but was significant different compared to that of W10 or 0.2% CPB A treatments (P<0.05). FCR of chicks treated with PBSL or other pellet binders tended to improve compared to that of control.

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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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    • v.26 no.2
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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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    • v.41 no.5
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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.

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

  • Jeon, Tae-Dong;Lee, Han-Gil;Hong, Seo-Yuong;Heo, Dong-Seok;Yoon, Il-Ji
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.1
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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
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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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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