• Journal of Animal Science and Technology
• /
• v.49 no.3
• /
• pp.351-358
• /
• 2007

This experiment was conducted to determine correlation between in vitro protein fractions and in situ protein degradation rate with major dairy protein sources(soybean meal, corn gluten meal, cotton seed meal, kapok seed meal and perilla meal). Five protein fractions were obtained according to the Cornell Net Carbohydate and Protein System(CNCPS), and in situ protein degradation rates were determined by technique using nylon bags incubated for 0, 4, 8, 12 and 24hrs in the rumen of three Holstein steers. Fraction A was highest in kapok seed meal(14.6%) and lowest in corn gluten meal(0.6%) (P<0.05). The highest B1, B2 and B3 fractions were contained in soybean meal(8.27%), cotton seed meal(74%), and perilla meal(40%), respectively. Corn gluten meal was very high in fraction C. In situ protein degradation rate of soybean meal was 98%, highest among five protein sources, and corn gluten meal had the lowest rate at 28%. Correlation analysis showed that easily soluble fractions of both methods, in situ protein degradation rate and digestible protein fractions, and in situ protein degradation rate minus “a” and fraction B2+B3 were highly correlated. These results indicate that in vitro protein fractionation can be used in the estimation of in situ protein degradation.

• Journal of Life Science
• /
• v.17 no.8 s.88
• /
• pp.1129-1134
• /
• 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.

• Korean Journal of Agricultural Science
• /
• v.44 no.3
• /
• pp.392-399
• /
• 2017

The present study was conducted to investigate the effects of different dietary proteins as fraction-enriched protein, defined by Cornell net carbohydrates and protein system (CNCPS), on in vivo ruminal fermentation pattern and blood metabolites in Holstein steers fed total mixed ration (TMR) containing 17.2% crude protein. Four ruminally cannulated Holstein steers in a $4{\times}4$ Latin square design consumed TMR only (control) and TMR with rapeseed meal (AB1), soybean meal (B2), and perilla meal (B3C). Each protein was substituted for 23.0% of crude protein in TMR. Rumen digesta were taken through ruminal cannula at 1 h interval during the feeding cycle in order to analyze ruminal pH, ammonia-N, and volatile fatty acids (VFA). Plasma metabolites in blood taken via the jugular vein after the rumen digesta sampling were analyzed. Feeding perilla meal significantly (p < 0.05) decreased mean ruminal pH compared with control and the other protein feeding groups. Compared with control, feeding protein significantly (p < 0.05) increased ruminal ammonia-N concentration except for AB1. Statistically (p > 0.05) similar total VFA appeared among control and the supplemented groups. However, control, AB1, and B2 showed higher (p < 0.05) acetate concentrations than B3C, and propionate was vice versa. CNCPS fractionated protein significantly (p < 0.05) affected concentrations of albumin and total protein in blood; i.e. plasma albumin was lower for control and B2 groups than AB1 and B3C groups. Despite lack of significances (p > 0.05) in creatinine and blood urea nitrogen, AB1 and B2 groups were numerically higher than the others.

• Journal of Life Science
• /
• v.28 no.9
• /
• pp.1007-1015
• /
• 2018

The E6-associated protein (E6AP) is known to induce the ubiquitination and proteasomal degradation of HCV core protein and thereby directly impair capsid assembly, resulting in a decline in HCV replication. To counteract this anti-viral host defense system, HCV core protein has evolved a strategy to inhibit E6AP expression via DNA methylation. In the present study, we further explored the mechanism by which HCV core protein inhibits E6AP expression. HCV core protein upregulated both the protein levels and enzyme activities of DNA methyltransferase 1 (DNMT1), DNMT3a, and DNMT3b to inhibit E6AP expression via promoter hypermethylation in HepG2 cells but not in Hep3B cells, which do not express p53. Interestingly, p53 overexpression alone in Hep3B cells was sufficient to activate DNMTs in the absence of HCV core protein and thereby inhibit E6AP expression via promoter hypermethylation. In addition, upregulation of p53 was absolutely required for the HCV core protein to inhibit E6AP expression via promoter hypermethylation, as evidenced by both p53 knockdown and ectopic expression experiments. Accordingly, levels of the ubiquitinated forms of HCV core protein were lower in HepG2 cells than in Hep3B cells. Based on these observations, we conclude that HCV core protein evades ubiquitin-dependent proteasomal degradation in a p53-dependent manner.

• Bulletin of the Korean Chemical Society
• /
• v.33 no.9
• /
• pp.3071-3075
• /
• 2012

Human ${\alpha}$-synuclein is the major component of the protein aggregates known as Lewy bodies or Lewy neurites, which define the intracellular lesions of Parkinson's disease. Despite extensive efforts, the physiological function of ${\alpha}$-synuclein has not yet been elucidated in detail. As an approach to defining its function, proteins that interacted with ${\alpha}$-synuclein were screened in phage display assays. The SNARE protein vesicle t-SNARE-interacting protein homologous 1B (VTI1B) was identified as an interacting partner. A selective interaction between ${\alpha}$-synuclein and VTI1B was confirmed by coimmunoprecipitation and GST pull-down assays. VTI1B and ${\alpha}$-synuclein were colocalized in N2a neuronal cells, and overexpression of ${\alpha}$-synuclein changed the subcellular localization of VTI1B to be more dispersed throughout the cytosol. Considering the role played by VTI1B, ${\alpha}$-synuclein is likely to modulate vesicle trafficking by interacting with a SNARE complex.

• Biomolecules & Therapeutics
• /
• v.12 no.3
• /
• pp.179-184
• /
• 2004

The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause for urinary tract infections. In our previous study, the Adhesin/CTXA2B was expressed as soluble recombinant chimaeric protein derived from the uropathogenic Escherichia coli adhesin genetically coupled to cholera toxin A2B (CTXA2B) subunit in Escherichia coli. Since it is very important to optimize IPTG concentration and culture temperature to maximize cell growth and productivity, These optimal culture factors were determined to increase the productivity of the expressed Adhesin/CTXA2B chimaeric protein in Escherichia coli TB1 carrying pMALfimH/ctxa2b. Our data demonstrate that optimal concentration of IPTG for increased production of chimaeric protein was 0.5 mM. Additionally, culture time was 10 hours and temperature, 37${\circ}C$.

• Tuberculosis and Respiratory Diseases
• /
• v.53 no.6
• /
• pp.595-611
• /
• 2002

Akt/PKB protein kinase B, ALI acute lung injury, ARDS acute respiratory distress syndrome, CREB C-AMP response element binding protein, ERK extracelluar signal-related kinase, fMLP fMet-Leu-Phe, G-CSF granulocyte colony-stimulating factor, IL interleukin, ILK integrin-linked kinase, JNK Jun N-terminal kinase, LPS lipopolysaccharide, MAP mitogen-activated protein, MEK MAP/ERK kinase, MIP-2 macrophage inflammatory protein-2, MMP matrix metalloproteinase, MPO myeloperoxidase, NADPH nicotinamide adenine dinucleotide phosphate, NE neutrophil elastase, NF-kB nuclear factor-kappa B, NOS nitric oxide synthase, p38 MAPK p38 mitogen activated protein kinase, PAF platelet activating factor, PAKs P21-activated kinases, PMN polymorphonuclear leukocytes, PI3-K phosphatidylinositol 3-kinase, PyK proline-rich tyrosine kinase, ROS reactive oxygen species, TNF-${\alpha}$ tumor necrosis factor-a.

• Journal of Dairy Science and Biotechnology
• /
• v.26 no.1
• /
• pp.5-10
• /
• 2008

This study was conducted to isolate protein components from culture supernatant of Lactobacillus casei. and measure anti-cancer activity. The protein components were isolated A and B on Ultrafiltration membrane(3, 10, 30, 100 KDa). And the protein components A and B were isolated fractions(number $3{\sim}9$) on FPLC. Experimental studies were progressed through the cell cytotoxicity and anti-cancer activities. Cell cytotoxicity test using human kidney normal cell(293) showed cytotoxicity of below 20% by the protein components A and B($100{\mu}g/mL$). The anti-cancer activity was increased up to 70% by the protein components A and B($100{\mu}g/mL$) in AGS(stomach cancer), A549(lung cancer), MCF-7 (breast cancer), SK-OV-3(ovary cancer) and LoVo(colon cancer). Cell cytotoxicity test was showed cytotoxicity of about 50% by the fractions(number 3, 8, 9) isolated FPLC. The others have not the cytotoxicity about the human normal cell. The anti-cancer activity was increased up to 70% by the fraction number 7 in cancer cell line. Therefore the components isolated from culture supernatant of Lactobacillus casei were showed anti-cancer activity.

• Korean Journal of Pharmacognosy
• /
• v.35 no.1 s.136
• /
• pp.16-21
• /
• 2004

Protein tyrosine phosphatase 1B(PTP1B) is thought to be a negative regulator in insulin signal-transduction pathway. Insulin-resistance by the activation of PTP1B is a hallmark of both type 2 diabetes and obesity. Thus, the compounds inhibiting PTP1B can improve insulin resistance and can be effective in treating type 2 diabetes and obesity. The methanol extracts of 160 herbal medicines were screened for the inhibitory activity against PTP1B. Among the tested extracts, methanol extracts of Amsonia elliptica, Areca catechu, Benincasa hispida, Morus alba, Salvia miltiorrhiza, Siegesbeckia orientalis, and Trichosanthes kirilowii showed relatively strong inhibitory activity against PTP1B.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.21 no.2
• /
• pp.151-155
• /
• 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.