• Journal of Microbiology and Biotechnology
• /
• v.27 no.12
• /
• pp.2156-2164
• /
• 2017

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

• Journal of Life Science
• /
• v.21 no.6
• /
• pp.907-911
• /
• 2011

Using silkworm Bombyx mori Bm5 cells, we established a stable cell line expressing the human keratinocyte growth factor (hKGF), named by the Bm5-hKGF cell, in which the protein hKGF is synthesized in the cell and secreted in the cell culture supernatant (CCS) at approximately 15-20 ng/ml. When the Bm5-hKGF cell was co-expressed with B. mori protein disulfide isomerase (bPDI) cDNA, its secretion increased by about two times the original amount. Through wound healing migration assay, it was demonstrated that the secreted hKGF included in the CCS has a very powerful biological activity of keratinocyte proliferation. We expect to produce useful human recombinant proteins from silkworm cultured cells in large quantities at low prices.

• Journal of Life Science
• /
• v.24 no.7
• /
• pp.743-749
• /
• 2014

The aim of this study was to identify the effect of fermented Angelica gigas Nakai (A. gigas) on lipid metabolism in orotic acid-induced fatty liver model rats. Sprague-Dawley male rats were randomly divided into four dietary groups (n=6 per group): a normal (N) group fed a standard diet only, OA control, OA acid plus 5% (w/w) A. gigas (OAG), and OA plus 5% (w/w) fermented A. gigas (OFAG). OA treatment induced enlargement of the liver and accumulation of hepatic triglycerides. The consum ption of fermented A. gigas reduced triglyceride concentrations in the liver and increased the serum lipid concentrations to normal levels. Furthermore, OA treatment significantly decreased serum triglyc eride concentrations without diminishing mRNA expression of microsomal triglyceride transfer protei n (MTP) and protein disulfide isomerase (PDI). Hepatic MTP mRNA expression increased 1.08-fold in response to OA treatment, despite triglyceride accumulation in the liver relative to that of the normal group. OFAG administration was slightly lower as compared to the OA treatment. This result suggests that MTP mRNA expression is not always correlated with hepatic triglyceride accumulation in the OA-induced fatty liver model. However, PDI mRNA expression was significantly increased in the OAG and OFAG groups (1.62-fold and 1.63-fold, respectively) compared with the normal group. The hepatocytes in the OA group contained numerous large fat droplets. These were slightly reduced in the OFAG group.

• Proceedings of the Korean Society of Sericultural Science Conference
• /
• 2001.04a
• /
• pp.30-30
• /
• 2001

No Abstract. See Full-text

• Proceedings of the Korean Society of Sericultural Science Conference
• /
• 2005.04a
• /
• pp.34-34
• /
• 2005

• Proceedings of the Korean Society of Applied Entomolohy Conference
• /
• 2005.05a
• /
• pp.147-147
• /
• 2005

• Proceedings of the Korean Society of Sericultural Science Conference
• /
• 2002.10a
• /
• pp.34-34
• /
• 2002

No Abstract, See Full Text

• Journal of Korean Neurosurgical Society
• /
• v.59 no.2
• /
• pp.106-116
• /
• 2016

Objective : Protein disulfide isomerase (PDI) acts as a chaperone on the cell surface, and it has been reported that PDI is associated with the tumor cell migration and invasion. The aims of this study are to investigate the anti-migration effect of bacitracin, which is an inhibitor of PDI, and the associated factor in this process. Methods : U87-MG glioma cells were treated with bacitracin in 1.25, 2.5, 3.75, and 5.0 mM concentrations. Western blot with caspase-3 was applied to evaluate the cytotoxicity of bacitracin. Adhesion, morphology, migration assays, and organotypic brain-slice culture were performed to evaluate the effect of bacitracin to the tumor cell. Western blot, PCR, and gelatin zymography were performed to investigate the associated factors. Thirty glioma tissues were collected following immunohistochemistry and Western blot. Results : Bacitracin showed a cytotoxicity in 3rd (p<0.05) and 4th (p<0.001) days, in 5.0 Mm concentration. The cell adhesion significantly decreased and the cells became a round shape after treated with bacitracin. The migration ability, the expression of phosphorylated focal adhesion kinase (p-FAK) and matrix metalloproteinase-2 (MMP-2) decreased in a bacitracin dose- and time-dependent manner. The U87-MG cells exhibited low-invasiveness in the 2.5 mM, compared with the untreated in organotypic brain-slice culture. PDI was expressed in the tumor margin, and significantly increased with histological glioma grades (p<0.001). Conclusion : Bacitracin, as a functional inhibitor of PDI, decreased the phosphorylated FAK and the secreted MMP-2, which are the downstream of integrin and play a major role in cell migration and invasion, might become one of the feasible therapeutic strategies for glioblastoma.

• BMB Reports
• /
• v.42 no.10
• /
• pp.661-666
• /
• 2009

Porcine pancreas development is not well studied at the molecular level despite being a therapeutic resource for diabetic patients. In this study, we investigated expression of lineage markers and performed proteomic analysis. Expression of the early lineage markers Pdx1 and Ptf1a was developmentally conserved between mice and pigs, whereas expression of the islet differentiation marker Pax4 was delayed in porcine compared with murine pancreas development. Proteomic analysis found that expression levels of chymotrypsinogen were down-regulated during porcine pancreas development while those of digestive enzymes like lipases, elastase and serine protease were up-regulated. In addition, specific isoforms of protein folding assistants such as protein disulfide isomerase and prefoldin were expressed at specific stages during the maturation of digestive enzymes. Taken together, these results show that development of the porcine pancreas is regulated by a concerted interplay of pancreas lineage marker proteins and other specified proteins, resulting in a functional endocrine and exocrine organ.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.8
• /
• pp.1364-1367
• /
• 2008

Oxidative stress damages all cellular constituents, and therefore, cell has to possess various defense mechanisms to cope. Saccharomyces cerevisiae, widely used as a model organism for studying cellular responses to oxidative stress, contains three glutathione peroxidase (Gpx) proteins. Among them, Gpx3 plays a major defense role against oxidative stress in S. cerevisiae. In this study, in order to identity the new interaction proteins of Gpx3, we carried out two-dimensional gel electrophoresis after immunoprecipitation (IP-2DE), and MALDI-TOF mass spectrometry. The results showed that several proteins including protein disulfide isomerase, glutaredoxin 2, and SSY protein 3 specifically interact with Gpx3. These findings led us to suggest the possibility that Gpx3, known as a redox sensor and ROS scavenger, has another functional role by interacting with several proteins with various cellular functions.