• Title/Summary/Keyword: Molecular Spectroscopy

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Molecular analysis of peptide toxins secreted by various Pseudomonas tolaasii strains (다양한 Pseudomonas tolaasii 균주에 의해 분비되는 펩티드 독소의 분석)

  • Yun, Yeong-Bae;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.63 no.4
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    • pp.387-392
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    • 2020
  • Pseudomonas tolaasii is a pathogen causing brown blotch disease in cultivated mushrooms. In previous study, various strains of P. tolaasii were isolated from the mushrooms with disease symptoms and they were further divided into Ptα, Ptβ, and Ptγ subtypes according to the 16S rRNA gene analysis. To investigate the secretion of peptide toxins, tolaasin and its analog peptides, culture extracts of Pt group strains were analyzed by gel permeation chromatography. Those of Ptα subtype strains contained two chromatographic peaks, band A and B. Meanwhile, those of Ptβ and Ptγ subtype strains contained mainly band A component and a little of band B. Molecular weights of toxic peptides of culture extracts were measured by MALDI-TOF mass spectrometry. In Ptα subtype strains, the peptide compositions of band A and B were same including tolaasin I (1,987 Da), tolaasin II (1,943 Da), and its two analog peptides, 1,973 Da and 2,005 Da. The strains of Ptβ and Ptγ subtype secreted many components of MW 1,100-1,200 Da, but they did not synthesize any tolaasin-like peptides. These results suggest that the only Ptα subtype strains secrete tolaasin and its analog peptide toxins and the strains of Ptβ and Ptγ subtypes have different pathogenic characters causing brown blotch disease.

Preparation of a Hydrophobized Chitosan Oligosaccharide for Application as an Efficient Gene Carrier

  • Son Sohee;Chae Su Young;Choi Changyong;Kim Myung-Yul;Ngugen Vu Giang;Jang Mi-Kyeong;Nah Jae-Woon;Kweon Jung Keoo
    • Macromolecular Research
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    • v.12 no.6
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    • pp.573-580
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    • 2004
  • To prepare chitosan-based polymeric amphiphiles that can form nanosized core-shell structures (nanopar­ticles) in aqueous milieu, chitosan oligosaccharides (COSs) were modified chemically with hydrophobic cholesterol groups. The physicochemical properties of the hydrophobized COSs (COSCs) were investigated by using dynamic light scattering and fluorescence spectroscopy. The feasibility of applying the COSCs to biomedical applications was investigated by introducing them into a gene delivery system. The COSCs formed nanosized self-aggregates in aqueous environments. Furthermore, the physicochemical properties of the COSC nanoparticles were closely related to the molecular weights of the COSs and the number of hydrophobic groups per COS chain. The critical aggregation concentration values decreased upon increasing the hydrophobicity of the COSCs. The COSCs effi­ciently condensed plasmid DNA into nanosized ion-complexes, in contrast to the effect of the unmodified COSs. An investigation of gene condensation, performed using a gel retardation assay, revealed that $COS6(M_n=6,040 Da)$ containing $5\%$ of cholesteryl chloroformate (COS6C5) formed a stable DNA complex at a COS6C5/DNA weight ratio of 2. In contrast, COS6, the unmodified COS, failed to form a stable COS/DNA complex even at an elevated weight ratio of 8. Furthermore, the COS6C5/DNA complex enhanced the in vitro transfection efficiency on Human embryonic kidney 293 cells by over 100 and 3 times those of COS6 and poly(L-lysine), respectively. Therefore, hydrophobized chitosan oligosaccharide can be considered as an efficient gene carrier for gene delivery systems.

Localization of the Membrane Interaction Sites of Pal-like Protein, HI0381 of Haemophilus influenzae

  • Kang, Su-Jin;Park, Sung Jean;Lee, Bong-Jin
    • Molecules and Cells
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    • v.26 no.2
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    • pp.206-211
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    • 2008
  • HI0381 of Haemophilus influenzae was investigated by circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy. HI0381 is a 153-residue peptidoglycan-associated outer membrane lipoprotein, and a part of the larger Tol/Pal network. Here, we report its backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments, and secondary structure predictions. About 97% of all of the $^1HN$, $^{15}N$, $^{13}CO$, $^{13}C{\alpha}$, and $^{13}C{\beta}$ resonances covering 131 non-proline residues of the 134 residue, mature protein, were clarified by sequential and specific assignments. CSI and TALOS analyses revealed that HI0381 contains five ${\alpha}$-helices and five ${\beta}$-strands. To characterize the structure of HI0381, the effects of pH and salt concentration were investigated by CD. In addition, the structural changes occurring when HI0381 was in a membranous environment were investigated by comparing its HSQC spectra and CD data in buffer and in DPC micelles; the results showed that helix ${\alpha}4$ and strand ${\beta}4$ became aligned with the membrane. We conclude that the conformation of HI0381 is affected by the membrane environment, implying that its folded state is directly related to its function.

Syntheses and Characterizations of Serine and Threonine Capped Water-Dispersible ZnS:Mn Nanocrystals and Comparison Study of Toxicity Effects on the growth of E. coli by the Methionine, Serine, Threonine, and Valine Capped ZnS:Mn Nanocrystals

  • Lim, Eun-Ju;Park, Sang-Hyun;Byun, Jong-Hoe;Hwang, Cheong-Soo
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1741-1747
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    • 2012
  • Water-dispersible ZnS:Mn nanocrystals were synthesized by capping the surface of the nanocrystals with conventional aminoacids ligands: serine and threonine. The aminoacids capped ZnS:Mn nanocrystal powders were characterized by XRD, HR-TEM, EDXS, ICP-AES and FT-IR spectroscopy. The optical properties were also measured by UV/Vis and solution photoluminescence (PL) spectroscopies in aqueous solvents. The solution PL spectra showed broad emission peaks around 600 nm with PL efficiencies of 9.7% (ZnS:Mn-Ser) and 15.4% (ZnS:Mn-Thr) respectively. The measured particle sizes for the aminoacid capped ZnS:Mn nanocrystals by HR-TEM images were about 3.0-4.0 nm, which were also supported by Debye-Scherrer calculations. In addition, cytotoxic effects of four aminoacids capped ZnS:Mn nanocrsystals over the growth of wild type E. coli were investigated. Although toxicity in the form of growth inhibition was observed with all the aminoacids capped ZnS:Mn nanocrystals at higher dose (1 mg/mL), ZnS:Mn-Met and ZnS:Mn-Thr appeared non-toxic at doses less than 100 ${\mu}g$/mL. Low biological toxicities were seen at doses less than 10 ${\mu}g$/ mL for all nanocrystals.

Chemical Bonding and Surface Electronic Structures of Pt3Co (111), Pt3Ni (111) Single Crystals

  • Kim, Yong-Su;Jeon, Sang-Ho;Bostwick, Aaron;Rotenberg, Eli;Ross, Philip N.;Stamenkovic, Vojislav R.;Markovic, Nenad M.;Noh, Tae-Won;Han, Seung-Wu;Mun, Bong-Jin Simon
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.08a
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    • pp.139-139
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    • 2012
  • With angle resolved photoemission spectroscopy (ARPES), the surface electronic band structures of Pt3Co (111) and Pt3Ni (111) single crystals are investigated, which allow to study the bonding interaction between chemically absorbed atomic oxygen and its surfaces. The d-band electrons of subsurface TM are separated from the direct chemical bonding with atomic oxygen. That is, the TM does not contribute to direct chemical bonding with oxygen. From the density functional theory (DFT) calculations, it is identified that the main origin of improved oxygen absorption property, i.e. softening of Pt-O bonding, is due to the suppression of Pt surface-states which is generated from change of interlayer potential, i.e. charge polarization, between Pt-top and TM-subsurface. Our results point out the critical roles of subsurface TM in modifying surface electronic structures, which in turn can be utilized to tune surface chemical properties.

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Contribution of Hydrophobic Interactions to HubWA Folding Reaction (소수성 상호작용이 HubWA 단백질의 폴딩 반응에 끼치는 영향)

  • Park, Soon-Ho
    • Journal of the Korean Chemical Society
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    • v.63 no.6
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    • pp.427-434
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    • 2019
  • The role of hydrophobic residues on protein folding reaction was studied by folding kinetics measurements in conjunction with protein engineering. The HubWA, which was derived from human ubiquitin by mutating the residues at 45 (Phe to Trp) and 26 (Val to Ala), was used as a mutational background. Fourteen hydrophobic residues were mutated to alanine. Among fourteen variants generated, only four variant proteins (V5A, I13A, V17A, and I36A) were suitable for folding study. The folding kinetics of these variants was measured by stopped-flow fluorescence spectroscopy. The folding kinetics of HubWA and V17A was observed to follow a three-state on-pathway mechanism. On the other hand, folding kinetics of V5A, I13A, and I36A was observed to follow a two-state mechanism. Based on these observations, transition of protein folding reaction from collision-diffusion mechanism to nucleation-condensation mechanism was discussed.

Cell-SELEX Based Identification of an RNA Aptamer for Escherichia coli and Its Use in Various Detection Formats

  • Dua, Pooja;Ren, Shuo;Lee, Sang Wook;Kim, Joon-Ki;Shin, Hye-su;Jeong, OK-Chan;Kim, Soyoun;Lee, Dong-Ki
    • Molecules and Cells
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    • v.39 no.11
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    • pp.807-813
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    • 2016
  • Escherichia coli are important indicator organisms, used routinely for the monitoring of water and food safety. For quick, sensitive and real-time detection of E. coli we developed a 2'F modified RNA aptamer Ec3, by Cell-SELEX. The 31 nucleotide truncated Ec3 demonstrated improved binding and low nano-molar affinity to E. coli. The aptamer developed by us out-performs the commercial antibody and aptamer used for E. coli detection. Ec3(31) aptamer based E. coli detection was done using three different detection formats and the assay sensitivities were determined. Conventional Ec3(31)-biotin-streptavidin magnetic separation could detect E. coli with a limit of detection of $1.3{\times}10^6CFU/ml$. Although, optical analytic technique, biolayer interferometry, did not improve the sensitivity of detection for whole cells, a very significant improvement in the detection was seen with the E. coli cell lysate ($5{\times}10^4CFU/ml$). Finally we developed Electrochemical Impedance Spectroscopy (EIS) gap capacitance biosensor that has detection limits of $2{\times}10^4CFU/mL$ of E. coli cells, without any labeling and signal amplification techniques. We believe that our developed method can step towards more complex and real sample application.

NMR Spectroscopic Analysis on the Chiral Recognition of Noradrenaline by β-Cyclodextrin ( β-CD) and Carboxymethyl- β-cyclodextrin (CM- β-CD)

  • Lee, Sang-Hoo;Yi, Dong-Heui;Jung, Seung-Ho
    • Bulletin of the Korean Chemical Society
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    • v.25 no.2
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    • pp.216-220
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    • 2004
  • ${\beta}$-CD and CM- ${\beta}$-CD as chiral NMR shift agents were used to resolve the enantiomers of noradrenaline (NA). The stoichiometry of each complex formed between the CDs and the enantiomers of NA was found to be 1 : 1 through the continuous variation plots. The binding constants (K) of the complexes were determined from $^1H$ NMR titration curves. This result indicated that both ${\beta}$-CD and CM- ${\beta}$-CD formed the complexes with the S(+)-NA more preferentially than its R(-)-enantiomer. The K values for the complexes with ${\beta}$-CD ($K_{S(+)}$ = 537 $M^{-1}$ and $K_{R(-)}$ = 516 $M^{-1}$ was larger than those with CM- ${\beta}$-CD ($K_{S(+)}$ = 435 $M^{-1}$ and $K_{R(-)}$ = 313 $M^{-1}$), however, enantioselectivity (${\alpha}$) of S(+)- and R(-)-NA to CM- ${\beta}$-CD ( ${\alpha}$ = 1.38) was larger than that to ${\beta}$-CD ( ${\alpha}$ = 1.04), indicating that CM- ${\beta}$-CD was the better chiral NMR solvating agents for the recognition of the enantiomers of NA. Two dimensional rotating frame nuclear Overhauser enhancement spectroscopy (ROESY) experiments were also performed to explain the binding properties in terms of spatial fitting of the NA molecule into the macrocyclic cavities.

EFFECT OF WATER CONTENT ON THE FLEXURAL STRENGTH DURING REFIRING IN DENIAL PORCELAIN (치과용 도재의 재소성 과정중 수분 함량이 강도에 미치는 영향)

  • Park Hye-Yang;Shim June-Sung;Lee Keun-Woo
    • The Journal of Korean Academy of Prosthodontics
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    • v.41 no.5
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    • pp.656-673
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    • 2003
  • Statement of problem : Long-term exposure of dental porcelain to saliva during temporary cementation of a porcelain-fused to metal (PFM) restoration could affect mechanical strength of dental porcelain if the restoration is refired. Purpose : This work was performed to verify the effect of water on the mechanical strength in aged dental porcelain. Material and method : 63 specimens(Vintage Metalbond opaque and opal powder) were distributed to three experimental groups ; non-water immersed control, immersed and pedried, and immersed and non-predired groups. The changes in flexural strength and fracture toughness after specimen refiring related to Fourier Transform Infrared (FT-IR) spectroscopy. Results : 1. The FT-IR reflectances assigned to molecular bonds of $H_2O$ were noted as significantly different between the first-fired group and three refired groups and between two water-immersed groups and control group after refiring(p<0.05). They were also significantly different between predried group and non-predried group after refiring(p<0.05) 2. For opal specimens, FT-IR absorbances for hydrogen bond of $H_2O$ and silanols were significantly higher in non-predried group than in predreid group(p<0.05). 3 Predried opal group showed the highest mean flexural strength(p<0.05). Non-predried group indicated higher mean flexural strength than control group(p<0.05). 4. The mean fracture toughness for predired group was higher than non-predried group(p<0.05). 5. The difference of leucite crystal size is noted between control group and water-immersed, predried group in scanning electron microscopic study(${\times}10000$).

A SIGNATURE OF CHROMOSPHERIC ACTIVITY IN BROWN DWARFS: A RECENT RESULT FROM NIRLT MISSION PROGRAM

  • Sorahana, Satoko;Suzuki, Takeru K.;Yamamura, Issei
    • Publications of The Korean Astronomical Society
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    • v.32 no.1
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    • pp.131-133
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    • 2017
  • We present the latest results from the Mission Program NIRLT (PI: I.Yamamura), the near-infrared spectroscopy of brown dwarfs using the AKARI/IRC grism mode with the spectral resolution of ~ 120. The near-infrared spectra in the wavelength range between 2.5 and $5.0{\mu}m$ are especially important to study the brown dwarf atmospheres because of the presence of major molecular bands, including $CH_4$ at $3.3{\mu}m$, $CO_2$ at $4.2{\mu}m$, CO at $4.6{\mu}m$, and $H_2O$ around $2.7{\mu}m$. We observed 27 sources, and obtained 16 good spectra. Our model fitting reveals deviations between theoretical model and observed spectra in this wavelength range, which may be attributed to the physical condition of the upper atmosphere. The deviations indicate additional heating, which we hypothesize to be due to chromospheric activity. We test this effect by modifying the brown dwarf atmosphere model to artificially increase the temperature of the upper atmosphere, and compare the revised model with observed spectra of early- to mid-L type objects with $H{\alpha}$ emission. We find that the chemical structure of the atmosphere changes dramatically, and the heating model spectra of early-type brown dwarfs can be considerably improved to match the observed spectra. Our result suggests that chromospheric activity is essential to understand early-type brown dwarf atmospheres.