• Nuclear Engineering and Technology
• /
• v.52 no.10
• /
• pp.2320-2327
• /
• 2020

The extreme hydrophobicity of expanded polytetrafluoroethylene (ePTFE) hinders bone-tissue integration, thus limiting the use of ePTFE in medical implant applications. To improve the potential of ePTFE as a biomaterial, 2-hydroxyethyl methacrylate (HEMA) was grafted onto the ePTFE surface using the gamma irradiation technique. The characteristics of the grafted ePTFE were successfully evaluated using attenuated total reflectance Fourier transform infrared (ATR-FTIR), field-emission scanning electron microscopy (FESEM)/energy dispersive X-ray (EDX), and X-ray photoelectron spectroscopy (XPS). Under the tensile test, the modified ePTFE was found to be more brittle and rigid than the untreated sample. In addition, the grafted ePTFE was less hydrophobic with a higher percentage of water uptake compared to the untreated ePTFE. The protein adsorption test showed that grafted ePTFE could adsorb protein, which was denoted by the presence of N peaks in the XPS analysis. Moreover, the formation of the globular mineral on the grafted ePTFE surface was successfully visualized using the FESEM analysis, with a ratio of 1.94 for Ca:P minerals by the EDX. To summarize, the capability of the modified ePTFE to show protein adsorption and mineralization indicates the improvement of the polymer properties, and it can potentially be used as a biomaterial for implant application.

• Biomolecules & Therapeutics
• /
• v.30 no.6
• /
• pp.593-602
• /
• 2022

The human papillomavirus (HPV)-18 E7 (E7) oncoprotein is a major transforming protein that is thought to be involved in the development of cervical cancer. It is well-known that E7 stimulates tumour development by inactivating pRb. However, this alone cannot explain the various characteristics acquired by HPV infection. Therefore, we examined other molecules that could help explain the acquired cancer properties during E7-induced cancer development. Using the yeast two-hybrid (Y2H) method, we found that the Elk-1 factor, which is crucial for cell proliferation, invasion, cell survival, anti-apoptotic activity, and cancer development, binds to the E7. By determining which part of E7 binds to which domain of Elk-1 using the Y2H method, it was found that CR2 and CR3 of the E7 and parts 1-206, including the ETS-DNA domain of Elk-1, interact with each other. As a result of their interaction, the transcriptional activity of Elk-1 was increased, thereby increasing the expression of target genes EGR-1, c-fos, and E2F. Additionally, the colony forming assay revealed that overexpression of Elk-1 and E7 promotes C33A cell proliferation. We expect that the discovery of a novel E7 function as an Elk-1 activator could help explain whether the E7 has novel oncogenic activities in addition to p53 inactivation. We also expect that it will offer new methods for developing improved strategies for cervical cancer treatment.

• BMB Reports
• /
• v.36 no.5
• /
• pp.493-498
• /
• 2003

The scFv antibody towards the Burkholderia pseudomallei exotoxin was previously constructed by phage display and exhibited good specificity towards the exotoxin. We report here the optimization of the scFv expression in an E. coli expression system. Four different E. coli strains (ER2537, TG1, HB2151, and XL1-Blue) were examined for optimal expression of the scFv protein. Two types of carbon source (i.e. 0.2% glucose and 0.2% glycerol) were also tested for their ability to induce the scFv expression. Cells that carried the scFv construct were grown at $30^{\circ}C$ and induced with 0.05 mM IPTG. The expression was then monitored by SDS-PAGE, Western blotting, and indirect ELISA. The Western blot profile showed different levels of the scFv expression among the host strains; XL1-Blue exhibited the highest level of the scFv protein expression. Glycerol at a concentration of 0.2% (v/v) significantly increased the scFv protein expression level when compared to 0.2% (w/v) glucose. Further optimization demonstrated that the scFv protein expression in XL1-Blue was the most optimal with a glycerol concentration as low as 0.05%. However, by indirect ELISA, only the scFv protein that was expressed in 0.2% (v/v) glycerol exhibited high specificity towards the Burkholderia pseudomallei exotoxin.

• The Plant Pathology Journal
• /
• v.21 no.4
• /
• pp.334-342
• /
• 2005

The prototype Chlorella virus PBCV-l encodes 11 tRNA genes and over 350 protein-encoding genes in its 330 kbp genome. Initial attempts to overexpress the recombinant A189/192R protein, a putative virus attachment protein, in E. coli strain BL21(DE3) SI were unsuccessful, and multiple protein bands were detected on Western blots. However, the full-length A189/192R recombinant protein or fragments derived from it were detected when they were expressed in E. coli BL21 CodonPlus (DE3) RIL, which contains extra tRNAs. Codon usage analysis of the a189/192r gene showed highly biased usage of the AGA and AVA codons compared to genes encoded by E. coli and Chlorella. In addition, there were biases of XXA/U($56\%$) and XXG/ C($44\%$) in the codons recognized by the viral tRNAs, which correspond to the codon usage bias in the PBCV-1 genome of XXA/U ($63\%$) over those ending in XXC/G ($37\%$). Analysis of the codon usage in the major capsid protein and DNA polymerase showed preferential usage of codons that can be recognized by the viral tRNAs. The Asn (AAC) and Lys (AAG) codons whose corresponding tRNA genes are duplicated in the tRNA gene cluster were the most abundant (i.e., preferred) codons in these two proteins. The tRNA genes encoded in the PBCV-l genome seem to play a very important role during the synthesis of viral proteins through supplementing the tRNAs that are frequently used in viral proteins, but are rare in the host cells. In addition, these tRNAs would help the virus to adapt to a wide range of hosts by providing tRNAs that are rare in the host cells.

• Journal of Life Science
• /
• v.12 no.2
• /
• pp.164-172
• /
• 2002

Breast cancer cell lines display a wide variety of growth factor receptors, and considerable evidences implicate the importance of signalings from those receptors. A useful prognostic indicator would be the level of activity of a second messenger protein used in common by these receptors. Our studies were designed to obtain preliminary information on the possible role of polyamine as a mediator of the membrane-associated protein phosphorylation and as a regulator of second messenger in mitogenic signal of estrogen or growth factors in MCF-7 human breast lancer cells. DFMO significantly inhibited the phosphorylation induced by $E_2$, TGF-$\alpha$ and EGF in membrane-associated proteins (154, 134, 116, and 104 kDa). Exogenous polyamines abolished the inhibitory effect of DFMO. Tyrosine phosphorylations of membrane-associated proteins were not increased by $E_2$ or growth factor treatments and not affected DFMO treatment. Polyamine administration markedly enhanced the tyrosine phosphorylation of membrane-associated proteins (154, 134, and 116 kDa). In the present study, $E_2$ and TGF-$\alpha$ and EGF enhanced protein phosphorylation in the almost same levels. These data indicate that $E_2$ and growth factor signaling pathway may cross-talk through various protein kinase which phosphorylated many substrate proteins (154, 134, 116 and 104 kDa). Polyamines may be involved in growth signaling pathway of $E_2$ and TGF-$\alpha$ or EGF for the cross-talk through regulation of the protein phosphorylation such as 154, 134, 116 and 104 kDa. Polyamine may also selectively interfere with several different protein kinases, and the specific steps in signal transduction system were effected by polyamines.

• Proceedings of the Microbiological Society of Korea Conference
• /
• 1999.04a
• /
• pp.51.1-51.1
• /
• 1999

• Journal of Fisheries and Marine Sciences Education
• /
• v.26 no.4
• /
• pp.868-876
• /
• 2014

This study was conducted to estimate the optimum dietary protein to energy (P/E) ratio in juvenile river puffer. Nine experimental diets were formulated with three energy levels and three protein levels at each energy levels. Three energy levels of 3000, 3750 and 4500 kcal diets were included at 45, 50 and 55% crude protein (CP) levels, respectively (45P3000, 45P3750, 45P4500, 50P3000, 50P3750, 50P4500, 55P3000, 55P3750 and 55P4500). Fish averaging $3.43{\pm}0.02$ g randomly were fed the experimental diets in triplicate groups for 8 weeks. Weight gain of fish fed the 50P4500 diet were significantly higher than that of fish fed the 45P3000, 45P3750, 45P4500, 50P3000 and 50P3750 diets (P<0.05), but there was not significantly different from that of fish fed the 50P4500, 55P3750 and 55P4500 diets. Feed efficiency of fish fed the 50P4500 diet were significantly higher than that of fish fed the 45P3000, 45P3750, 45P4500, 50P3000 and 50P3750 diets (P<0.05), but there was not significantly different from that of fish fed the 50P4500, 55P3000, 55P3750 and 55P4500 diets. Protein efficiency ratio of fish fed the 45P3000 and 45P3750 diets was higher than that of fish fed 50P4500 and 55P4500, but there was not significantly different from that of fish fed the 45P3000, 45P3750, 45P4500, 50P3000, 50P3750, 55P3000 and 55P3750 diets. Based on weight gain, feed efficiency and specific growth rate, diets containing energy levels 4500 kcal/kg diet had an optimum P/E ratio of approximately 111 mg protein/kcal (50% crude protein) in juvenile river puffer.

• Biomolecules & Therapeutics
• /
• v.4 no.1
• /
• pp.97-102
• /
• 1996

To investigate whether human $\beta$$_2$-adrenergic receptor devoid of the C-terminal two transmembrane helices retain its ligand binding activity and specificity, 5'780-bp DNA fragment of the receptor gene which encodes amino acid 1-260 of human $\beta$$_2$-adrenergic receptor was subcloned into the bacterial fusion protein expression vector and expressed as a form of glutathione-S-transferase (GST) fusion protein in E. coli DH5$\alpha$. The receptor fusion protein was expressed as a membrane bound form which was verified by SDS-PAGE and Western blot. The fusion protein expressed in this study specifically bound $\beta$-adrenergic receptor ligand [$^3$H] Dihydroalprenolol. In saturation ligand binding assay, the $K_{d}$ value was 7.6 nM which was similar to that of intact $\beta$$_2$-adrenergic receptor in normal animal tissue ( $K_{d}$=1~2 nM) and the $B_{max}$ value was 266 fmol/mg membrane protein. In competition binding assay, the order of binding affinity of various adrenergic receptor agonists to the fusion protein was isoproterenol》epinephrine norepinephrine, which was similar to that of intact receptor in normal animal tissue. These results suggest that N-terminal five transmembrane helices of the $\beta$$_2$-adrenergic receptor be sufficient to determine the ligand binding activity and specificity, irrespective of the presence or absence of the C-terminal two transmembrane helices.s.s.s.

• The Korean Journal of Zoology
• /
• v.31 no.3
• /
• pp.157-164
• /
• 1988

SCK tumor cells were exposed to heat shock or several sulihydryl-reacting agents such as iodoacetamide(IAA), zinc chloride(Zn), and 2-mercaptoethanol(ME). Stress proteins induced by these agents were examined and the relationship between the induction of stress proteins and the production of abnormal proteins was discussed. Based on the present experiments, two classes of intracellular pathways for the induction of stress proteins were defined; one dependent on and the other independent of protein synthesis. The presence of cycloheximide during the induction period blocked the formation of stress proteins in the cells exposed to Zn or ME, but not in those exposed to heat shock or IAA.Therefore, stress protein seems to be induced either by denaturation of pre-existing mature proteins (e.g., heat shock or IAA) or by newly synthesized abnormal proteins(e.g., Zn or ME). In conclusion, it is ilkely that the production of abnormal proteins by stresses triggers stress protein induction. In addition, it was found that the cells exposed to IISP and GRP inducers simultaneously responded to more strong stress among several stresses encountered.

• Korean Journal of Food Science and Technology
• /
• v.2 no.2
• /
• pp.8-16
• /
• 1970

Exploitation of leaf protein concentrates for human consumption is very important. Leaf protein concentrates can be easily prepared by mechanically mincing leaves material and press it for getting the juice. Crude protein can be separated from the juice by aging, adjusting the pH, or heating to $75-80^{\circ}C$ etc. This report deals with the extractability of total-N from 69 species of fresh leaves by mechanical process, and then compared the recovery of leaf protein concentrates from leaf extracts by treating with TCA, pH adjustment and heating. Results are summarized as follows. 1. In general, the greater the content of total-N of leaves the greater the percentage extraction. Extraction of the juice from leaves is needed at least two times. The simple equations are constituted between the total-N (T; %) and the first and second extractability ($E_1,\;E_2;\;%$) of the total-N of leaves, as follows: $E_1=0.8168T\;E_2=0.1830T$ 2. The optimum pH value for coagulating protein from extracts is considered to be 3.5 to 4.5. However, the products of leaf protein concentrate by the pH adjustment of extracts are generally dull in color with rich elasticity. 3. Recoveries of the leaf protein concentrate from extracts by treating methods were in the following order of TCA treatment> pH 4 treatment> pH 3 treatment> heat treatment. The yield of leaf protein concentrates decreased bout 10% with pH 4 treatment, 11.4% with pH 3 treatment, and 14.8% with heat treatment compared with the TCA treatment. 4. The heat treatment is the most benifitial method for the production of leaf protein concentrates with regard to properties of texture, color and yield of products and easiness of the treatment method.