• Journal of Microbiology
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• v.42 no.3
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• pp.248-252
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• 2004

In order to identify the protein(s) secreted into culture medium by the sool-l/retl-l mutation of Saccharomyces cerevisiae, proteins from the culture medium of cells grown at permissive (28$^{\circ}C$) and non-permissive temperatures (37$^{\circ}C$), were analyzed. Comparison of protein bands separated by SDS-PAGE identified a prominent band of 47-kDa band from a mutant grown at 37$^{\circ}C$. N-terminal amino acid sequencing of this 47-kDa protein showed high identity with enolases 1 and 2. Western blot analysis revealed that most of the cell wall-bound enolase was released into the culture medium of the mutant grown at 37$^{\circ}C$, some of which were separated as those with lower molecular weights. Our results, presented here, indicate the impairment of cell wall enolase biogenesis and assembly by the sool-l/retl-l mutation of S. cerevisiae.

• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.212-215
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• 2009

The inhibitory effects of Curcuma longa rhizome-derived materials against nitric oxide (NO) production were assessed. The inhibitory effect (57%) on NO production was evidenced by the methanol extract of C. longa at $1\;{\mu}g/mL$. In the fractionation of the methanol extract, the ethyl acetate fraction evidenced an inhibitory effect greater than 62.1% at $1\;{\mu}g/mL$. The active constituent was identified as curcumin. Curcumin exerted potent inhibitory effects of 78.7 and 65.7% at concentrations of 1 and $0.5\;{\mu}g/mL$, respectively. Furthermore, the inhibitory effect of ar-turmerone was measured as 31.3 and 15.8% at 1 and $0.5\;{\mu}g/mL$, respectively. The iNOS expression-suppressive effects of curcumin were assessed via western blot analysis. Our results suggest that curcumin and ar-turmerone may prove useful in the development of new types of NO inhibitors.

• Biomedical Science Letters
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• v.19 no.3
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• pp.224-232
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• 2013

Cucurbitacins are a natural cell-permeable triterpenoid compound isolated from Cucurbitaceae and Cruciferae. Cucurbitacins have been used as folk medicine because of their anti-inflammatory and analgesic effects. In the present study, we investigate the anti-cancer effects of cucurbitacin I on colitis-associated colon carcinogenesis induced by azoxymethane (AOM)/dextran sodium sulfate (DSS) in BALB/c mice. Cucurbitacin I treatment attenuated loss of body weight and decreased the number of colon tumors. Western blot analysis showed that cucurbitacin I treatment significantly inhibited the protein expression of inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6. These results suggest that cucurbitacin I suppressed inflammatory reaction and tumor development in colitis-associated colon carcinogenesis.

• Korean Journal Plant Pathology
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• v.14 no.3
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• pp.203-208
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• 1998

We isolated and characterized an antifungal peptide from the seeds of Phytolacca americana. Growth inhibition assay with Botrytis cinerea was used to screen inhibitory proteins from 60 different plant species. A 4 kDa antifungal peptide (Pa-AFP) inhibitory to hyphal growth of B. cinerea was found in the seeds of P. americana. The peptide Pa-AFP was purified to homogeneity by chromatographies of Sephadex G-50, DEAE-Sepharose, Sephacryl S-300, and C18 reverse-phase HPLC. Western blot analysis showed that a polyclonal antibody raised against the purified peptide cross-reacted with a 4 kDa protein in seeds but not in root and leaf tissues of P. americana. Pa-AFP inhibited the hyphal growth of Botrytis cinerea, Rihzoctonia solani, Fusarium oxysporum, and Magnaporthe grisea. Pa-AFP exhibited growth inhibition of Saccharomyces cerevisiae strain BWG7a, which was sensitive to osmotin.

• Journal of Plant Biotechnology
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• v.36 no.3
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• pp.224-229
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• 2009

Human lactoferrin is an iron-binding glycoprotein with many biological activities, including the protection against microbial and virus infection and stimulation of the immune system. We introduced a human lactoferrin (hLf) cDNA under the control of 35S promoter into sweet potato by particle bombardment. Transgenic plants were regenerated via somatic embryogenesis. Transgenic plants were produced typical tuberous roots in soil. PCR, Southern and northern analyses confirmed that the hLf cDNA was incorporated into the plant genome and was properly expressed in plants. Western blot analysis showed that the 80 kDa full length hLf protein was produced in transgenic tuberous roots. Overall results indicated that sweet potato would be an excellent host to produce human therapeutic proteins.

• International Journal of Industrial Entomology and Biomaterials
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• v.34 no.1
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• pp.1-5
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• 2017

Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

• Journal of the Korean Magnetic Resonance Society
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• v.12 no.1
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• pp.33-39
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• 2008

Four sesterterpenoids were isolated from AMPK activity-guided fraction of marine sponge Sarcotragus species. Their planar structures were determined from combination of extensive 1D and 2D NMR experiments and MS data, and the configuration at the chiral centers were assigned by comparison with the $^1H$ and $^{13}C$ chemical shifts of the known compounds. Among four compounds, compound 2 was found to be a new sarcotrine derivative. Though not strong, compounds 1-4 moderately showed AMPK activation effect on L6 myoblast cell through Western Blot analysis.

• Animal Bioscience
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• v.35 no.4
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• pp.624-630
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• 2022

Objective: The aim of this study was to investigate the expression level of filaggrin-2 (FLG2) in correlation with drip loss. Methods: The muscle samples were randomly taken from a local meat supplier. Samples were taken from Longissimus lumborum muscles to evaluate the drip loss (n = 100). Five muscles per group (low and high drip loss) were selected to evaluate FLG2 mRNA and protein expression levels. Results: mRNA of FLG2 gene was not significantly different in pigs with different levels of drip loss (p>0.05). Statistical analysis revealed that FLG2 protein expression levels were significantly different between the drip loss groups. Western blot revealed that the high drip loss group had higher FLG2 protein expression level than the low drip loss group (p<0.001). Moreover, immunohistochemistry revealed the high signal intensity was on the muscle cell membrane and cytoplasm. Conclusion: FLG2 protein might play roles in drip loss of pork and will provide the basis for information to improving meat quality traits in pigs.

• Journal of Veterinary Clinics
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• v.21 no.1
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• pp.20-22
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• 2004

Bacterial infection of canine atopic dermatitis is largely caused by Staphylococcus intermedius and may be a superficial or deep pyoderma. The Purpose of this study was to identify the major proteins of S. intermedius cell surface components in humoral immune response of atopic dermatitis dog. Sera samples were obtained from dogs with atopic dermatitis and superficial pyoderma referred to the Veterinary Medical Teaching Hospital, Konkuk University. An isolate of S. intermedius from a clinical case of canine atopic dermatitis was cultured in brain heart infusion broth overnight at $37^{\circ}C$ in aerobic conditions on an orbital shaker. Following culture, Staphylococci were harvested by centrifugation, washed in PBS, and resuspended in PBS containing lysostaphin. The soluble components were separated by centrifugation and were collected. The soluble extract of S. intermedius was separated by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were electrophoretically transferred onto nitrocellulose membrane. Western blotting for the specificity of serum IgG antistaphylococcal antibody was performed with anti-dog-IgG and sera obtained from an atopic dermatitis case and a normal dog. The molecular masses of four major proteins of S. intermedius recognized by serum obtained from an atopic dermatitis case were 18, 31, 75, and 110 kDa as determined by Western blot analysis. The present study indicates that most dogs of S. intermedius infection with atopic dermatitis could have a significant humoral immune response to bacterial proteins of the causative organism.

• Journal of Acupuncture Research
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• v.21 no.2
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• pp.41-55
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• 2004

Objective : To investigate the possible molecular mechanism (s) of melittin as a candidate of anti-cancer drug, we examined the effects of the compound on the growth of human lung carcinoma cell line A549. Methods : Growth inhibitory study, flow cytometry analysis, SDS-polyacrylamide gel electrophoresis and Western blot analysis, RT-PCR and in vitro caspases activity assay were performed. Results : Melittin treatment declined the cell viability of A549 cells in a concentration-dependent manner, which was associated with induction of apoptotic cell death. Melittin treatment down-regulated the levels of Bcl-XS/L mRNA and protein expression of A549 cells, an anti-apoptotic gene, however, the those of Bax, a pro-apoptotic gene, were up-regulated. Melittin induced the proteolytic cleavage and activation of caspase-3 and caspase-9 protease in a dose-dependent manner without alteration of inhibitor of apoptosis proteins family and Akt expression. Western blot analysis and RT-PCR data revealed that the levels of tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 were also remained unchanged. Conclusions : Taken together, these findings suggest that melittin-induced inhibition of human lung cancer cell growth is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and melittin may have therapeutic potential in human lung cancer.