• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1098-1105
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• 2007

Quantum mechanical and molecular dynamics simulation analysis has been performed on the model system for CALB (Candida antarctica lipase B) with esters to study the reaction mechanism and conformational preference of catalytic hydrolysis and the esterification reaction. Using quantum mechanical analysis, the ping-pong bi-bi mechanism was applied and energies and 3-dimensional binding configurations of the whole reaction pathways were calculated. Further molecular dynamics simulation analysis was performed on the basis of the transition state obtained from quantum mechanical study to observe the effect of structures of the substrates. Calculation results using substrates of different chain length and chiral configurations were compared for conformational preference. The calculated results showed very small influence on chain length, whereas chiral conformation showed big differences. Calculated results from molecular modeling studies have been compared qualitatively with the experimental data using racemic mixtures of (${\pm}$)-cis-4-acetamido-cyclopent-2-ene-1-ethyl acetate as substrates.

• Journal of the Korean Magnetic Resonance Society
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• v.14 no.2
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• pp.117-126
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• 2010

Hsp33 is a molecular chaperone achieving a holdase activity upon response to a dual stress by heat and oxidation. Despite several crystal structures available, the activation process is not clearly understood, because the structure inactive Hsp33 as its reduced, zinc-bound, monomeric form has not been solved yet. Thus, we initiated structural investigation of the reduced Hsp33 monomer by NMR. In this study, to overcome the high molecular weight (33 kDa), the protein was triply isotope-[$^{13}C$, $^{15}N$, $^2H$]-labeled and its inactive, monomeric state was ensured. 2D-[$^1H$, $^{15}N$]-TROSY and a series of triple resonance spectra could be successfully obtained on a high-field (900 MHz) NMR machine with a cryoprobe. However, under all of the different conditions tested, the number of resonances observed was significantly less than that expected from the amino acid sequence. Thus, a possible contribution of dynamic conformational exchange leading to a line broadening is suggested that might be important for activation process of Hsp33.

• KIEE International Transactions on Electrophysics and Applications
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• v.3C no.2
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• pp.59-65
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• 2003

Electrical properties such as permittivity and tan$\delta$ of unaged (control) and aged (72 h at 18$0^{\circ}C$) mica/epoxy composites of 130 ${\mu}{\textrm}{m}$ thickness were measured and their surface conditions were characterized using scanning electron microscopy (SEM), Fourier transform infrared (FTIR), electron spectroscopy for chemical analysis (ESCA), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). Both permittivity and tan6 of control specimens were higher than those of short-time aged specimens. FTIR results show a new peak at 1710 $cm^{-1}$ / for short-time aged specimens, originating from carbonyl group formed by the oxidation reaction during the aging process. ESCA results show that the binding energy at 532.9 eV representing the singlet state of oxygen ( $O_{1s}$) decreases by 13.7%, whereas that at 534.6 eV increases by 13.7%. Glass transition temperatures of control and short-time aged specimens are observed to be 95.4$^{\circ}C$ and 113.4$^{\circ}C$, which increase with the increase of aging time. TGA results indicate that the control specimens contain a smaller amount of volatile components than the short-time aged specimens.s.

• Journal of Microbiology
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• v.39 no.3
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• pp.186-194
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• 2001

Scrub typhus, caused by Orientia tsutsugamushi infection, is clinically and histopathologically characterized by local as well as systemic inflammatory reactions, indicating that orientiae induce mechanisms that amplify the inflammatory response. To reveal underlying mechanisms of chemoattraction and activation of responding leukocytes, expression of chemokine and tumor necrosis factor alpha (TNF-$\alpha$) genes in murine peritoneal macrophages after infection with the obligate intracellular bacterium Ο.tsutsugamushi was investigated. The genes that were unregulated included macrophage inflammatory proteins l$\alpha$/$\beta$(MIP-l$\alpha$/$\beta$), MIP-2, monocyte chemoattractant protein 1(MCP-1), RANTES (regulated upon activation, normal T-cell expressed and secreted), gamma-interferon-inducible protein 10(IP-10) and TNF-$\alpha$. Peak expression of these chemokines and TNF-$\alpha$ was observed between 1 and 3 h after infection. These responses returned to or approached baseline preinfection levels 6 h after challenge. Semiquantitative reverse transcription (RT)-PCR analysis revealed dramatic Increases during infection in the steady-state levels of mRNA ceding for the inhibitory subunit of NF-kB (IkB$\alpha$), whose transcription is enhanced by binding of NF-kB within the IkB$\alpha$promoter region. Thus, Ο. tsutsugamushi appears to be a stung inducer of chemokines and TNF-$\alpha$ which may significantly contribute to inflammation and tissue damage observed in scrub typhus by attracting and activating phagocytic leukocytes.

• Animal cells and systems
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• v.11 no.2
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• pp.169-173
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• 2007

Hyperpolarization-activated nonselective cation channels (HCNs) play a pivotal role in producing rhythmic electrical activity in the heart and the nerve cells. In our previous experiments, voltage-dependent $Cd^{2+}$ access to one of the substituted cysteines in S6, T464C, supports the existence of an intracellular voltage-dependent activation gate. Direct binding of intracellular cAMP to HCN channels also modulates gating. Here we attempted to locate the cAMP-modulated structure that can modify the gating of HCN channels. SpHCN channels, a sea urchin homologue of the HCN family, became inactivated rapidly and intracellular cAMP removed this inactivation, resulting in about eight-fold increase of steady-state current level. T464C was probed with $Cd^{2+}$ applied to the intracellular side of the channel. We found that access of $Cd^{2+}$ to T464C was strongly gated by cAMP as well as voltage. Release of bound $Cd^{2+}$ by DMPS was also gated in a cAMP-dependent manner. Our results suggest the existence of an intracellular cAMP-modulated gate in the lower S6 region of spHCN channels.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.3
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• pp.500-504
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• 2001

Gamma irradiation was applied to reduce egg allergy. Ovalbumin (OVA), an egg white protein, was used as model allergen and was gamma-irradiated at 3, 5, or 10 kGy in an aqueous state (2.0 mg/mL). The changes in allergenic and antigenic properties of OVA resulted from gamma irradiation were monitored by ELISA with serum from egg-hypersensitive patients (H-IgE), and mouse monoclonal IgG (M-IgG) or rabbit polyclonal IgG (R-IgG). The binding ability of H-IgE to irradiated OVA was dose-dependently reduced. However, IgGs from animal did better recognize 3 or 5 kGy-irradiated OVA. In the evaluation of immune reactivity using blind test, the reactivity of H-IgE rapidly decreased depending upon the irradiation dose. However, the reactivities of M-IgG and R-IgG was higher at 5 and 3 kGy-irradiated OVA than non-irradiated control. The results provide a new possibility to use irradiation process for reducing the allergenicity of egg white.

• Journal of the Korean Ceramic Society
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• v.41 no.2
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• pp.97-101
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• 2004

W $O_{x}$-based semiconductor type thin film gas sensor was fabricated for the detection of N $O_{x}$ by reactive d.c. sputtering method. The relative oxidation state of the deposited W $O_{x}$ films was approximately compared by the calculation of the difference of the binding energy between Ols to W4 $f_{7}$2/ core level XPS spectra in the standard W $O_3$ powder of known composition. As the annealing temperature increased from 500 to 80$0^{\circ}C$, relative oxygen contents and grain size of the sputtered films were gradually increased. As the results of sensitivity ( $R_{gas}$/ $R_{air}$) measurements for the 5 ppm N $O_2$ gas, the sensitivity was 110 and the sensor showed recovery time as fast as 200 s. The other sensor properties were examined in terms of surface microstructure, annealing temperature, and relative oxygen contents. These results indicated that the W $O_3$ thin film with well controlled structure is a good candidate for monitoring and controlling of automobile exhaust.haust.t.t.t.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 2010.06a
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• pp.376-376
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• 2010

The single crystals of p-$CdIn_2Te_4$ were grown by the Bridgman method without the seed crystal. From photocurrent measurements, it was found that three peaks, A, B, and C, correspond to the intrinsic transition from the valence band states of $\Gamma_7$(A), $\Gamma_6$(B), and $\Gamma_7$(C) to the conduction band state of $\Gamma_6$, respectively. The crystal field splitting and the spin orbit splitting were found to be 0.2360 and 0.1119 eV, respectively, from the photocurrent spectroscopy. The temperature dependence of the $CdIn_2Te_4$ band gap energy was given by the equation of $E_g(T)=E_g(0)-(9.43{\times}10^{-3})T^2/(2676+T)$. $E_g$(0) was estimated to be 1.4750, 1.7110, and 1.8229 eV at the valence band states of A, B, and C, respectively. The band gap energy of p-$CdIn_2Te_4$ at room temperature was determined to be 1.2023 eV.

• BMB Reports
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• v.51 no.10
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• pp.484-485
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• 2018

Receptor-interacting protein kinase-3 (RIP3 or RIPK3) is a serine-threonine kinase largely essential for necroptotic cell death; it also plays a role in some inflammatory diseases. High levels of RIP3 are likely sufficient to activate necroptotic and inflammatory pathways downstream of RIP3 in the absence of an upstream stimulus. For example, we have previously detected high levels or RIP3 in the skin of Toxic Epidermal Necrolysis patients; this correlates with increased phosphorylation of MLKL found in these patients. We have long surmised that there are molecular mechanisms to prevent anomalous activity of the RIP3 protein, and so prevent undesirable cell death and inflammatory effects when inappropriately activated. Recent discovery that Carboxyl terminus of Hsp 70-Interacting Protein (CHIP) could mediate ubiquitylation- and lysosome-dependent RIP3 degradation provides a potential protein that has this capacity. However, while screening for RIP3-binding proteins, we discovered that pellino E3 ubiquitin protein ligase 1 (PELI1) also interacts directly with RIP3 protein; further investigation in this study revealed that PELI1 also targets RIP3 for proteasome-dependent degradation. Interestingly, unlike CHIP, which targets RIP3 more generally, PELI1 preferentially targets kinase active RIP3 that has been phosphorylated on T182, subsequently leading to RIP3 degradation.

• Archives of Pharmacal Research
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• v.21 no.3
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• pp.291-297
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• 1998

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.