• Bulletin of the Korean Chemical Society
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• 제18권9호
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• pp.933-938
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• 1997

The interaction of mastoparan B, a tetradecapeptide toxin found in the hornet Vespa basalis, with phospholipid bilayers was investigated. Synthetic mastoparan B and its analogs, obtained by substituting one hydrophilic amino acid (2-Lys, 4-Lys, 5-Ser, 8-Ser, 11-Lys, or 12-Lys) in mastoparan B with Ala, were studied. Mastoparan B and its analogs were synthesized by the solid-phase method. As shown by circular dichroism spectra, mastoparan B and its analogs adopted an unordered structure in buffer solution. All peptides took an α-helical structure, and the α-helical content of its analogs increased in the presence of neutral and acidic liposomes as compared to that of mastoparan B. In the calcein leakage experiment, we observed that mastoparan B interacted more weakly with lipid bilayers in neutral and acidic media than its analogs. Mastoparan B also showed slightly lower antimicrobial activity and hemolytic activity towards human erythrocytes than its analogs. These results indicate that the greater hydrophobicity of the amphiphilic α-helix of mastoparan B by replacement with alamine residues results in the increased biological activity and helical content.

• Bulletin of the Korean Chemical Society
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• 제8권6호
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• pp.441-444
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• 1987

The circular dichroism(CD) spectra of the optically active [$Co(acac)_2(diamine)]^+$ complexes were measured in the several protic and aprotic solvents, where acac = acetylacetonate anion and diamine = ethylenediamine and trimethylenediamine. The degree of the CD variation in protic solvents was enhanced as the dielectric constant decreases except n-butanol and benzylalcohol. And the degree of the CD variation in aprotic solvents was roughly increased as both dipole moment and dielectric constant decrease except aromatic solvents and the solvents having no dipole moment. It was deduced that the CD variations of the complexes have been due to the conformational change of acetylacetonate ligands coordinated to Co(III) ion.

• Bulletin of the Korean Chemical Society
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• 제19권4호
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• pp.444-449
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• 1998

The kinetics of the formation of triplex $poly(dA){\cdot}[poly(dT)]_2$ from poly(dA)·poly(dT) and poly(dT) is examined by various optical spectroscopic methods, including absorption, circular and linear dichroism (LD) spectroscopy. In the pseudo first order condition, where the poly(dT) concentration is kept lower than that of duplex, the association of the poly(dT) is enhanced by the presence of ethidium; the rate constant is proportional to the amount of ethidium in the mixture. When the concentration of the duplex and the single strand is the same, a spectral change is explained by double exponential curves, indicating that at least two steps are involved, the fast association and slow rearrangement steps. In contrast to the pseudo first order kinetics, the association step in an equimolar condition is not affected by the presence of ethidium. In the rearrangement step, the magnitude of LD decreases with an increase in ethidium concentration, suggesting that the bending of polynucleotide around the intercalation site occurs in the rearrangement step.

• Bulletin of the Korean Chemical Society
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• 제18권5호
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• pp.510-514
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• 1997

We studied the interaction of 4',6-diamidino-2-phenylindole (DAPI) with single stranded poly(dA) using optical spectroscopic methods, including absorption, circular dichroism (CD), and fluorescence spectroscopy. The temperature-dependent conformation of poly(dA) was also investigated. The conformation of poly(dA) varied with temperature, which is explained by the stacking-destacking process of the adenine bases, resulting from the sugar conformation. The hypochromicity and red-shift in the absorption spectroscopy, the lack of CD change in the drag absorption region, and the fluorescence behavior, especially a great accessibility of the I2 quencher to the poly(dA)-bound DAPI, suggest that DAPI binds to the outside of poly(dA). The Job plot for the DAPI-poly(dA) mixture demonstrated that a stoichiometry of one DAPI molecule binds to the one phosphate of poly(dA).

• Bulletin of the Korean Chemical Society
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• 제20권9호
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• pp.1073-1077
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• 1999

CRAMP, a 37-amino acid cationic antimicrobial peptide was recently deduced from the cDNA cloned from mouse femoral marrow RNA. In order to investigate the structure-activity relationship and functional region of CRAMP, CRAMP and its 18-mer overlapping peptides were synthesized by the solid phase method. CRAMP showed broad spectrum antibacterial activity against both Gram-positive and Gram-negative bacterial strains (MIC: 3.125-6.25 μM) but had no hemolytic activity until 50 μM. CRAMP was found to have a potent anticancer activity (IC50: 12-23 μM) against two human small cell lung cancer cell lines. Furthermore, CRAMP was found to display faster bactericidal rate in B. subtilis rather than E. coli in the kinetics of bacterial killing. Among 18-meric overlapping fragment peptides, only CRAMP (16-33) displayed potent antibacterial activity (MIC: 12.5-50 μM) against several bacteria with no hemolytic activity. Circular dichroism (CD) spectra anal-ysis indicated that CRAMP and its analogues will form the amphipathic α-helical conformation in the cell membranes similar to other antimicrobial peptides, such as cecropins and magainins.

• 한국자기학회지
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• 제5권5호
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• pp.575-578
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• 1995

X-ray magnetic-circular-dichroism (MCD) spectra, orbital ($$) and spin magnetic moments ($$) for Co(110) monolayers a free standing mode or sandwiched between Pd(Pd/1Co/Pd)and Cu layers (Cu/1Co/Cu) are calculated using the thin film full potential linearized augmented plane wave energy band method. In contrast to the double peak structure predicted for the Co(0001) surface, only a minor side peak is found in the MCD spectra for Cu/Co/Cu, while MCD spectra for the other systems show a single peak structure. The MCD sum rules originally derived from a single ion model are found in the band approach to be valid for the systems investigated. However, for the spin sum rule, the magnetic dipole term ($$) is not negligible and needs to be included.

• Bulletin of the Korean Chemical Society
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• 제26권11호
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• pp.1728-1734
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• 2005

The interactions of Cu(II)-meso-Tetrakis(n-N-methylpyridiniumyl)porphyrin (n = 2,3,4), respectively referred to as o-, m- and p-CuTMPyP, and DNA, poly$[d(A-T)_2]$ and poly$[d(G-C)_2]$ were investigated by circular and linear dichroism (CD and LD). In the o-CuTMPyP case, in which the rotation of the pyridinium ring is prevented, the shape of the CD spectrum when associated to DNA and poly$[d(A-T)_2]$ resembles and is characterized by a positive band at a low drug to DNA concentration ratio (R ratio) and is bisignate at a high R ratio. The former CD spectrum shape has been attributed to porphyrin that is bound monomerically outside of DNA while the latter can be attributed to those that are stacked. When o-CuTMPyP is bound to poly$[d(G-C)_2]$, the excitonic CD appeared at a relatively high R ratio. In contrast, a characteristic negative CD band in the Soret region was apparent for both m- and p-CuTMPyP when bound to DNA and poly$[d(G-C)_2]$ at the low R ratios, indicating that the porphyrin molecule intercalates. However, the DNA is bent near the intercalation site and the plane of the porphyrin molecule tilts relative to the DNA helix axis, as judged by the magnitude of the reduced LD. Various stacking patterns were identified by the shape of the CD spectrum for m- and p-CuTMPyP when bound to poly$[d(A-T)_2]$. Three species for the former complex and two for the latter complex were found which may reflect the extent of the stacking.

• Journal of Microbiology and Biotechnology
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• 제24권6호
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• pp.748-755
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• 2014

In the genome annotation of Escherichia coli MG1655, the orf382 (1,149 bp) is designated as a gene encoding an alcohol dehydrogenase that may be Fe-dependent. In this study, the gene was amplified from the genome by PCR and overexpressed in Escherichia coli BL21(DE3). The recombinant $6{\times}$His-tag protein was then purified and characterized. In an enzymatic assay using different hydroxyl-containing substrates (n-butanol, $\small{L}$-threonine, ethanol, isopropanol, glucose, glycerol, $\small{L}$-serine, lactic acid, citric acid, methanol, or $\small{D}$-threonine), the enzyme showed the highest activity on $\small{L}$-threonine. Characterization of the mutant constructed using gene knockout of the orf382 also implied the function of the enzyme in the metabolism of $\small{L}$-threonine into glycine. Considering the presence of tested substrates in living E. coli cel ls and previous literature, we believed that the suitable nomenclature for the enzyme should be an $\small{L}$-threonine dehydrogenase (LTDH). When using $\small{L}$-threonine as the substrate, the enzyme exhibited the best catalytic performance at $39^{\circ}C$ and pH 9.8 with $NAD^+$ as the cofactor. The determination of the Km values towards $\small{L}$-threonine (Km = $11.29{\mu}M$), ethanol ($222.5{\mu}M$), and n-butanol ($8.02{\mu}M$) also confirmed the enzyme as an LTDH. Furthermore, the LTDH was shown to be an ion-containing protein based on inductively coupled plasma-atomic emission spectrometry with an isoelectronic point of pH 5.4. Moreover, a circular dichroism analysis revealed that the metal ion was structurally and enzymatically essential, as its deprivation remarkably changed the ${\alpha}$-helix percentage (from 12.6% to 6.3%).

• BMB Reports
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• 제40권6호
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• pp.911-920
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• 2007

Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.

• 한국진공학회:학술대회논문집
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• 한국진공학회 1999년도 제17회 학술발표회 논문개요집
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• pp.169-169
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• 1999

We studied magnetic properties of co thin films deposited on Pd (111) surface, which attracted much attention recently due to the perpendicular magnetic anisotropy, using magnetic circular dichroism(MCD). Special attention was paid on the effect of Pd capping and interface roughness on the direction of magnetic easy axis, and for that purpose MCD signals for all Co thicknesses were measured with two different ways : in-plane and out-of-plane geometry. In case of bare Co films deposited on smooth Pd(111) surface, no MCD signal was observed under 4$\AA$ co thickness. At 4$\AA$ Co thickness, MCD signal at the out-of-plane geometry was observed, and for thicker Co films, only in-plane MCD signal was measured. This type of magnetic easy axis transition has been reported for other cases like Co/Pt system. The effect of 5$\AA$ Pd capping on these bare Co films made an remarkable change on the transition of magnetic easy axis. Out-of-plane MCD signals exists up to 20$\AA$ Co thickness, and disappears at 24$\AA$ Co thickness. In-plane MCD signals first appears at 10$\AA$ Co thickness and gradually increases up to 24$\AA$ Co thickness. Between 10$\AA$ and 20$\AA$ Co thickness, in-plane and out-of-plane MCD signal coexist. The formation of multi-domain structure or the existence of tilted magnetic easy axis is an possible scenario for such an interesting coexistence. The effect of interface roughness was also tested by measuring MCD signal on Co films deposited on un-annealed Pd(111) surface. Out-of-plane MCD signal was observed up to 8$\AA$ Co thickness and the anisotropy of MCD signal at 4$\AA$ Co thickness was very large with respect to that of Co film deposited on the smooth substrate. Above 8$\AA$ thickness, there exists only in-plane MCD signal. From above results, it was concluded that both Pd capping and interface roughness induce and reenforce the perpendicular magnetic anisotropy. The large perpendicular magnetic anisotropy of Co/Pd multilayer system made by sputtering method can be well understood from our results.