• Journal of Life Science
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• v.13 no.4
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• pp.541-550
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• 2003

E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.

• BMB Reports
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• v.43 no.2
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• pp.140-145
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• 2010

The present study investigated whether the mammalian target of rapamycin (mTOR) signal pathway is involved in the regulation of high glucose-induced intramuscular adipogenesis in porcine muscle satellite cells. High glucose (25 mM) dramatically increased intracellular lipid accumulation in cells during the 10-day adipogenic differentiation period. The expressions of CCAAT/enhancer binding protein-$\alpha$ (C/EBP-$\alpha$) and fatty acid synthase (FAS) protein were gradually enhanced during the 10-day duration while mTOR phosphorylation and sterol-regulatory- element-binding protein (SREBP)-1c protein were induced on day 4. Moreover, inhibition of mTOR activity by rapamycin resulted in a reduction of SREBP-1c protein expression and adipogenesis in cells. Collectively, our findings suggest that the adipogenic differentiation of porcine muscle satellite cells and a succeeding extensive adipogenesis, which is triggered by high glucose, is initiated by the mTOR signal pathway through the activation of SREBP-1c protein. This process is previously uncharacterized and suggests a cellular mechanism may be involved in ectopic lipid deposition in skeletal muscle during type 2 diabetes.

• Journal of Life Science
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• v.8 no.4
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• pp.400-409
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• 1998

Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.6
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• pp.763-770
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• 1993

To investigate the quality changes during frozen storage, top shell, Omphalius pfeifferi capenteri, was stored at -18$^{\circ}C$, -$25^{\circ}C$ and -3$0^{\circ}C$ immediately after shelling and water holding capacity, protein composition and histological features were examined with the lapsed period of the storage. During the storage period, amount of free drip was increased with higher frozen temperature and longer frozen period, but with the longer storage period, the lower water holding capacity was observed. The extractability and composition of muscle protein, sarcoplasmic protein and stroma protein were rather stable regardless of frozen temperature and frozen storage period. However, the extractability of myofibrillar protein was decreased with higher frozen temperature and longer frozen storage period. On the changes of muscle tissue structure, following points were observed. 1) In the muscle tissue structure of fresh sample, fine muscle fiber was closely distributed all over the tissue regardless of cross and longitudinal section. 2) In tissue structure under frozen state, it was observed that ice crystals apparently grew with the higher storage temperature. Empty spaces between muscle bundles which wee formed by aggregations of muscle fiber were observed after 3 months storage at -18$^{\circ}C$ . 3) Tissue structure in thawed state was restored satisfactorily after 1 month storage regardless of storage temperature. After 3 months storage at -3$0^{\circ}C$, muscle tissue was well restored, but at -18$^{\circ}C$, empty spaces were apparent due to incomplete restoration.

• Archives of Plastic Surgery
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• v.34 no.1
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• pp.13-17
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• 2007

Purpose: To understand the pathogenesis of the disease that presents abnormally proliferated vascular endothelial cells, a model of DMH(1,2-dimethylhydrazine)-induced abnormal proliferation of HUVECs(Human Umbilical Vein Endothelial Cells) was made. We indirectly determined that Protein Kinase C(PKC) restricts the cellular proliferation and inhibits the manifestation of growth factor by using several inhibiting substances of the transmitter through our previous studies. Thereupon, we attempted to observe direct enzymatic activities of PKC and its correlation with the abnormal proliferation of vascular endothelial cells. Methods: $10^5$ HUVECs cells were applied to 6 individual well plates in three different groups; A control group cultured without treatment, a group concentrated with $0.75{\times}10^{-8}M$ DMH only, and a group treated with DMH & $5{\times}10^{-9}M$ Calphostin C, inhibitor of PKC. In analyzing the formation of intracellular PKC enzyme, protein separation was performed, and separated protein was quantitatively measured. PKC enzyme reaction was analyzed through Protein Kinase C Assay System (Promega, USA), and the results were analyzed according to Beer's law. Results: Enzymatic activity of PKC presented the highest in all reaction time of a group concentrated only with DMH, and the lowest in the control group. The group treated with DMH and the inhibitor revealed statistically lower enzymatic activity than group only with DMH in all reaction time, although higher than the control group. Conclusion: From the enzymatic aspect, most active and immediate reaction of the PKC was observed in the group concentrated with DMH only. The group treated with DMH & PKC inhibitor showed meaningful decrease. Accordingly, PKC holds a significant role in DMH-induced abnormal proliferation of vascular endothelial cells.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.1
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• pp.35-42
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• 1998

Whole faba beans (WFB) were dry roasted at different temperatures (110, 130, $150^{\circ}C$) for 15, 30, 45 minutes to determine the optimal heating conditions of time and temperature to increase nutritional value. Ruminant degradation characteristics of crude protein (CP) of WFB were determined by the nylon bag incubation technique in dairy cows fed 60% hay and 40% concentrate. Measured characteristics of crude protein (CP) were soluble (washable) fraction (S), undegradable fraction (U), lag time (T0), potentially degradable fraction (D) and the rate of degradation (Kd) of insoluble but degradable fraction. Based on measured characteristics, percentage bypass crude protein (%BCP) and bypass crude protein (BCP in g/kg) were calculated. Degradability of CP was reduced by dry roasting (p < 0.01). S was reduced rapidly with increasing time and temperature, from 49.0% in the raw WFB (RWFB) to 26.3% in $150^{\circ}C/45$ min. D varied from 50.7% in RWFB to 73.7% in $150^{\circ}C/45^{\prime}$. U varied from 0% in $130^{\circ}C/45^{\prime}$, $150^{\circ}/30^{\prime}$ and $150^{\circ}/45^{\prime}$ to 0.66% in $110^{\circ}/45^{\prime}$ (0.24% for the RWFB). Lag time (T0) varied from 1.58 h in $130^{\circ}C/30^{\prime}$ to 2.40 h in $150^{\circ}C/45^{\prime}$ (1.87 h for RWFB). Kd varied from 24.2% in the $110^{\circ}C/30^{\prime}$ to 4.3% in $150^{\circ}C/45^{\prime}$ (21.4% for the RWFB). Kd was significantly reduced with time and temperature. All these effects resulted in increasing % BCP from 8.9% in the $110^{\circ}C/45^{\prime}$, 11.3% in the RWFB to 43.1% in the $150^{\circ}C/45$. Therefore BCP increased from 31.3 and 39.9 to 148.4 g/kg respectively. Both %BCP and BCP at $150^{\circ}C/45$ increased nearly 4 times over the raw faba beans. The effects of dry roasting temperature and time on %BCP and BCP seemed to be linear up to the highest values tested. Therefore no optimal dry roasting conditions of time and temperature could be determined at this stage. It was concluded that dry roasting was effective in shifting crude protein degradation from rumen to intestine to reduce unnecessary nitrogen (N) loss in the rumen. To determine the optimal treatment, the digestibility of each treatment should be measured in the next trial using mobile bags technique.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.10
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• pp.1668-1675
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• 2004

The effect of pH on surface hydrophobicity, sulfhydryl group, infrared spectrum, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) pattern and enthalpy was investigated in recovered protein from mackerel and frozen blackspotted croaker by alkaline processing. Hydrophobic residue in myofibrillar protein exposed to the surface of protein, and hydrophobic interaction were the highest around 6$0^{\circ}C$. The surface hydrophobicity was different between myofibrillar protein and myofibrillar protein including sarcoplasmic protein (recovered protein). The peak at 1636 c $m^{-l}$ was increased with pH, and the recovered protein was unfolded in alkali pH. Difference of surface and total sulfhydryl group at pH 7.0 and 10 was comparative high, and decrease of surface sulfhydryl group indicated formation of S-S bonds. Mackerel and frozen blackspotted croaker in alkaline pH showed bands of polymerized myosin heavy chain on SDS-PAGE pattern. The transition temperatures of recovered protein were 33.1, 44.3 and 65.5$^{\circ}C$. Gelation of recovered protein from alkali processing was estimated by increase of $\beta$-sheet structure by pH treatment, S-S bonds by oxidation of surface sulfhydryl group in heating, polymerization of myosin heavy chain in order.r.

• Molecules and Cells
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• v.24 no.3
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• pp.437-440
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• 2007

$OGL-20P^T$-358 is a novel 66 amino acid residue protein from the hyperthermophile Thermococcus thioreducens sp. nov., strain $OGL-20P^T$, which was collected from the wall of the hydrothermal black smoker in the Rainbow Vent along the mid-Atlantic ridge. This protein, which has no detectable sequence homology with proteins or domains of known function, has a calculated pI of 4.76 and a molecular mass of 8.2 kDa. We report here the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of $OGL-20P^T$-358. Assignments are 97.5% (316/324) complete. Chemical shift index was used to determine the secondary structure of the protein, which appears to consist of primarily ${\alpha}$-helical regions. This work is the foundation for future studies to determine the three-dimensional solution structure of the protein.

• Applied Biological Chemistry
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• v.35 no.6
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• pp.501-506
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• 1992

Pepsin-catalyzed hydrolysis and plastein reaction were carried out to prepare plastein product from soymilk residue protein. Conditions required for optimal hydrolysis of soymilk residue protein and subsequent plastein production were investigated. The optimum substrate concentration, enzyme-substrate ratio, pH, reaction temperature and incubation time for hydrolysis were 3%, 1/50, 1.7, $45^{\circ}C$ and 24 hours, respectively. Plastein formation from peptic hydrolysate of soymilk residue protein was most effective at substrate concentratin of 40%, pH 4 and $45^{\circ}C$. Reaction time of 18 hours and enzyme-substrate ratio of 1/100 were selected for plastein production. Electrophoresis of the products revealed that protein-like substances of high molecular weight were produced from the plastein reaction.

• Journal of Nutrition and Health
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• v.30 no.6
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• pp.614-621
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• 1997

The experiments were performed to investigate the effects of protein and protein hydrolysates on lipid metabolism in the hyperlipidemic/hypercholesterolemic rat model induced by feeding fat-enriched diet. In Except 1 male rats were fed four semi-purified high fat and cholesterol diets that contained different nitrogen source, casein(C), casein hydrolysate(CH), corn gluten(G) and corn gluten hydrolysate(GH), for 6 weeks. In Expt. 2 rats were fed high fat and cholesterol diet for 4 weeks to induce hyperlipidemia and hypercholesterolemia. Then the rats were divided into 4 groups and were fed the four kinds of above experimental diets for 4 weeks consecutively. The contents of total lipid , cholesterol and triglyceride in blood, liver and feces were determined. Serum lipid concentrations of CH, G and GH were significantly lower than that of C. Serum cholesterol concentrations of hydrolysate groups(CH and GH) were significantly lower than those of intact protein groups(C and G). Serum HDL -cholesterol concentration tended to increase by hydrolysate intake. The total lipid, cholesterol contents in liver showed similarity results as above. Fecal lipid excretions of CH, G, and GH groups were significantly higher than that of C group. These results indicate that hypolipidemic and /or hypocholesterolemic effect of corn gluten or protein hydrolysates were detected in the process of inducing hyperlipidemia by high-fat and cholesterol diet or after inducing hyperlipidemia.