• BMB Reports
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• v.41 no.1
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• pp.62-67
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• 2008

The present study was aimed to elucidate the mechanism of stabilization of tubulin by deuterium oxide ($D_2O$). Rate of decrease of tryptophan fluorescence during aging of tubulin at 4$^{\circ}C$ and 37$^{\circ}C$ was significantly lower in $D_2O$ than in $H_2O$. Circular dichroism spectra of tubulin after incubation at 4$^{\circ}C$, suggested that complete stabilization of the secondary structure in D2O during the first 24 hours of incubation. The number of available cysteine measured by DTNB reaction was decreased to a lesser extent in $D_2O$ than in $H_2O$. . During the increase in temperature of tubulin, the rate of decrease of fluorescence at 335 nm and change of CD value at 222 nm was lesser in $D_2O$. Differential Scanning calorimetric experiments showed that the $T_m$ values for tubulin unfolding in $D_2O$ were 58.6$^{\circ}C$ and 62.17$^{\circ}C$, and in $H_2O$. those values were 55.4$^{\circ}C$ and 59.35$^{\circ}C$.

• Korean Chemical Engineering Research
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• v.44 no.1
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• pp.92-96
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• 2006

Enzymes in organic media afford many advantages such as chiral synthesis and resolution, modification of fats and oils and production of biodegradable polymers. However, the nature of solvents influences the activity and stability of enzymes, and the presence of organic solvents always constitute a risk of enzyme inactivation. Heat-shock protein Hsp90, one of the molecular chaperone, was applied for understanding of enzyme inactivation and for increasing of enzyme stability in water-miscible organic solvent. Hsp90 showed stabilization effect on HRP in the 30％ of DMSO, in the 30％ and 50％ of dioxane. Hsp90 also showed reactivation effect on the inactivated HRP by water-miscible organic solvent such as dioxane and DMSO. In addition, structural analysis using fluorescence spectrophotometry and circular dichroism showed that exposure of HRP in water-miscible organic solvent caused appreciable conformational changes and enzyme inactivation, and the unfolded HRP by water-miscible organic solvent was refolded by Hsp90.

• Korean Journal of Food Science and Technology
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• v.31 no.6
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• pp.1488-1494
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• 1999

Effect of ${\gamma}-irradiation$ on the SDS-PAGE pattern, secondary structure content, the solubility of commercial soy protein isolate (SPI) and whey protein concentrate (WPC) was investigated. The change in the subunit molecular weight of SPI and WPC irradiated in aqueous solution or dried state was studied using SDS-PAGE. The SDS-PAGE pattern of SPI irradiated in aqueous solution revealed the fragmentation and aggregation of the subunit protein. For WPC irradiated in aqueous solution. fragmentation of the subunit protein up to 10 kGy was observed. In contrast, ${\gamma}-irradiation$ of SPI and WPC in dried state did not cause any significant changes in the SDS-PAGE pattern. The change In the secondary structure of irradiated SPI and WPC solution was studied using circular dichroism. The aperiodic structure content of SPI and WPC solution increased at higher irradiation doses, which suggests that ${\gamma}-irradiation$ caused the disruption of the ordered structure of SPI and WPC solution. Gamma-irradiation also caused the change in solubility of SPI and WPC in dried state.

• Journal of the Korean Magnetic Resonance Society
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• v.20 no.3
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• pp.95-101
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• 2016

Apolipoprotein B-100 (Apo-B100) is a major component of low density lipoprotein (LDL). Apo B-100 protein has 4,536 amino acid sequence and these amino acids are classified into peptide groups A to G with subsequent 20 amino acids (P1-P302). The peptide groups were act as immunoglobulin (Ig) antigens which oxidized via malondialdehyde (MDA). The mimetic peptide P1 (EEEMLENVSLVCPKDAT RFK) out of D-group peptides carrying the highest value of IgG antigens were selected for structural studies that may provide antigen specificity. Circular Dichroism (CD) spectra were measured for peptide secondary structure in the range of 190-250 nm. Experimental results show that P1 exhibit partial of ${\beta}-sheet$ and random coil structure. Homonuclear (COSY, TOCSY, NOESY) 2D-NMR experiments were carried out for NMR signal assignments and structure determination for P1. On the basis of these completely assigned NMR spectra and distance data, distance geometry (DG) and Molecular dynamics (MD) were carried out to determine the structures of P1. The proposed structure was selected by comparisons between experimental NOE spectra and back calculated 2D NOE results from determined structure showing acceptable agreement. The total Root-Mean-Square-Deviation (RMSD) value of P1 obtained upon superposition of all atoms was in the range $0.33{\AA}$. The solution state P1 has mixed structure of ${\beta}-sheet$ (Glu[1] to Cys[12]) and random coil (Pro[13] to Lys[20]). These NMR results are well consistent with secondary structure from experimental results of circular dichroism. Structural studies based on NMR may contribute to the studies of atherosclerosis and observed conformational characteristics of apo B-100 in LDL using monoclonal antibodies.

• Journal of the Korean Magnetic Resonance Society
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• v.24 no.4
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• pp.136-142
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• 2020

Apolipoprotein B-100 (apo B-100), the main protein component that makes up LDL (Low density lipoprotein), consists of 4,536 amino acids and serves to combine with the LDL receptor. The oxidized LDL peptides by malondialdehyde (MDA) or acetylation in vivo were act as immunoglobulin (Ig) antigens and peptide groups were classified into 7 peptide groups with subsequent 20 amino acids (P1-P302). The biomimetic peptide P99 (KGTYG LSCQR DPNTG RLNGE) out of B-group peptides carrying the highest value of IgM antigens were selected for structural studies that may provide antigen specificity. Circular Dichroism (CD) spectra were measured for peptide secondary structure in the range of 190-260 nm. Experimental results show that P99 has pseudo α-helice and random coil structure. Homonuclear (COSY, TOCSY, NOESY) 2D-NMR experiments were carried out for NMR signal assignments and structure determination for P99. On the basis of these completely assigned NMR spectra and proton distance information, distance geometry (DG) and molecular dynamic (MD) were carried out to determine the structures of P99. The proposed structure was selected by comparisons between experimental NOE spectra and back-calculated 2D NOE results from determined structure showing acceptable agreement. The total Root-Mean-Square-Deviation (RMSD) value of P99 obtained upon superposition of all atoms were in the set range. The solution state P99 has mixed structure of pseudo α-helix and β-turn(Gln[9] to Thr[13]). These NMR results are well consistent with secondary structure from experimental results of circular dichroism. Structural studies based on NMR may contribute to the prevent oxidation studies of atherosclerosis and observed conformational characteristics of apo B-100 in LDL using monoclonal antibodies.

• Macromolecular Research
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• v.12 no.3
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• pp.276-281
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• 2004

In this study we investigated the spectral properties, including electric absorption, circular and linear dichroism (CD and LD), and fluorescence emission, of DNA in a DNA-branched polyethyleneimine (BPEI) polyplex at various polymer molecular weights (M$\_$w/) and BPEI-amine-to-DNA-phosphate ratios (N/P ratios). All BPEIs exhibited a common N/P dependence in their absorption and CD spectra. At N/P ratios ＜ 1.0, we observed some hyperchromism in the absorption spectrum, red-shifts in CD bands, and decreases in LD intensity and fluorescence intensity of intercalated ethidium. At intermediate N/P ratios, complete collapse of all spectra occurred. As the N/P ratio increased further, the polyplex dissolved in water. From its characteristic CD spectrum obtained under these conditions, we conclude that the DNA exists in a B-like form. The fluorescence and LD intensities never recovered even at high N/P ratios- which indicates that the dissolved polyplex possesses positive charges and the DNA in the polyplex is condensed despite its B-form CD spectrum. The N/P range in which the absorption and CD signals collapsed was wider when the BPEIs M$\_$w/ decreased. In the case where the BPEIs M$\_$w/ was 0.8 k, recovery of the absorption and CD spectral properties at a high N/P ratio was never achieved, which suggests that the molecular weight of the polymer plays an important role in its dissolution at a high N/P ratio.

• Bulletin of the Korean Chemical Society
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• v.32 no.11
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• pp.4083-4085
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• 2011

• Proceedings of the Korean Society of Dyers and Finishers Conference
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• 2011.03a
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• pp.35-35
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• 2011

The design and preparation of novel dye rotaxanes have gained much interest recently, since such structure usually exhibits peculiar spectral and optical changes. In spite of the promising results to date, increasing pressure remains to develop novel supramolecular structures based on stimuli-responsive systems. This presentation covers the study of inclusion complexes of cyclodextrins and various chromophores, with an emphasis on our most recent outcome of anisotropic hydrogel. In this system, physical gelation prepared from simple mixture of CD and a azo dye is completed through specific host-guest interaction. The obtained hydrogel exhibits respective morphological transitions based on supramolecular assembly and dissociation, leading to either precipitation or a sol-to-gel transition. It can identify different classes of metal ions, and, among them, naked-eye differentiation of lead ion is possible due to the coordination-induced unthreading of dye molecules. Accompanying structural changes were verified by numerous characterization techniques, including 2D-ROESY, HR-MAS, UV-Visible absorption, small-angle X-ray scattering, and induced circular dichroism measurements. Such properties discussed here will find useful in analytical applications, such as metal ion sensing and removal applications.

• Journal of the Korean Magnetic Resonance Society
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• v.20 no.1
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• pp.17-21
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• 2016

Periostin, a component of extracellular matrix (ECM) protein, is produced and secreted by the fibroblasts that are involved in chronic allergic inflammation diseases and various types of human cancers. Periostin protein is composed of multiple domains including four FAS1 domains which play important roles in cell adhesion and tumor metastasis by interacting with integrins. In spite of their important biological role, the structural information of periosin FAS1 domains was not revealed yet. Recently we systemically prepared various constructs of the FAS1 domains and tried to express them in E. coli. Of them, only single FAS1-II and -IV domains were highly soluble. Circular dichroism (CD) and nuclear magnetic resonance (NMR) studies revealed that the FAS1-IV domain might be suitable for three-dimensional structure determination using NMR spectroscopy.

• Bulletin of the Korean Chemical Society
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• v.16 no.1
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• pp.42-47
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• 1995

Interpolymer complex formation between basic polypeptide poly(L-proline) Form Ⅱ (PLP(Ⅱ)) and acidic polypeptides poly(L-glutamic acid) (PLGA) and poly(L-aspartic acid)(PLAA) has been studied in water-methanol (1:2 v/v) mixed-solvent by viscometry, potentiometry, light scattering and circular dichroism (CD) measurements. It has been found that polymer complexes between PLP(Ⅱ) and PLGA (or PLAA) are formed via hydrogen bonding with a stoichiometric ratio of PLP(Ⅱ)/PLGA (or PLAA)=1:2 (in unit mole ratio) and that PLP(Ⅱ) forms polymer complex more favorably with PLGA than with PLAA. In addition, the minimum (for pH 5.0) and the maximum (for pH 3.2) in reduced viscosity of dilute PLP(Ⅱ)-PLGA mixed solutions are observed at 0.67 unit mole fraction of PLGA (i.e., [PLP(Ⅱ)]/[PLGA]=1/2). These findings could be explained in terms of molecular structure (or conformation) of the complementary polymers associated with the complex formation.