• Title/Summary/Keyword: Intrinsic kinetic

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In vitro Folding of Recombinant Hepatitis B Virus X-Protein Produced in Escherichia coli: Formation of Folding Intermediates

  • Kim, Sun-Ok;Sohn, Mi-Jin;Jeong, Soon-Seog;Shin, Jeh-Hoon;Lee, Young-Ik
    • BMB Reports
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    • v.32 no.6
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    • pp.521-528
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    • 1999
  • The folding of recombinant hepatitis B virus X-protein (rHBx) solubilized from Escherichia coli inclusion bodies was investigated. By sequential dialysis of urea, rHBx was folded into its native structure, which was demonstrated by the efficacy of its transcriptional activation of the adenovirus major late promoter (MLP), fluorescence spectroscopy, and circular dichroism (CD) analysis. The decrease in CD values at 220 nm and a corresponding blue shift of the intrinsic fluorescence emission confirmed the ability of rHBx to refold in lower concentrations of urea, yielding the active protein. Equilibrium and kinetic studies of the refolding of rHBx were carried out by tryptophan fluorescence measurements. From the biphasic nature of the fluorescence curves, the existence of stable intermediate states in the renaturation process was inferred. Reverse phase-high performance liquid chromatography (RP-HPLC) analysis further demonstrated the existence of these intermediates and their apparent compactness.

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Band Alignment at CdS/wide-band-gap Cu(In,Ga)Se2 Hetero-junction by using PES/IPES

  • Kong, Sok-Hyun;Kima, Kyung-Hwan
    • Transactions on Electrical and Electronic Materials
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    • v.6 no.5
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    • pp.229-232
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    • 2005
  • Direct characterization of band alignment at chemical bath deposition $(CBD)-CdS/Cu_{0.93}(In_{1-x}Ga_x)Se_2$ has been carried out by photoemission spectroscopy (PES) and inverse photoemission spectroscopy (IPES). Ar ion beam etching at the condition of the low ion kinetic energy of 400 eV yields a removal of surface contamination as well as successful development of intrinsic feature of each layer and the interfaces. Especially interior regions of the wide gap CIGS layers with a band gap of $1.4\~1.6\;eV$ were successfully exposed. IPES spectra revealed that conduction band offset (CBO) at the interface region over the wide gap CIGS of x = 0.60 and 0.75 was negative, where the conduction band minimum of CdS was lower than that of CIGS. It was also observed that an energy spacing between conduction band minimum (CBM) of CdS layer and valance band maximum (VBM) of $Cu_{0.93}(In_{0.25}Ga_{0.75})Se_2$ layer at interface region was no wider than that of the interface over the $Cu_{0.93}(In_{0.60}Ga_{0.40})Se_2$ layer.

Identification of an Essential Tryptophan Residue Residue in Alliinase from Garlic (Allium sativum) by Chemical Modification

  • Jin, Yeong Nam;Choe, Yong Hun;Yang, Cheol Hak
    • Bulletin of the Korean Chemical Society
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    • v.22 no.1
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    • pp.68-76
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    • 2001
  • We have employed chemical modification to identify amino acids essential for the catalytic activity of alliinase (EC 4.4.1.4) from garlic (Allium sativum). Alliinase degrades S-alkyl-L cysteine sulfoxides, causing the characteristic odor of garlic. The activity of alliinase was rapidly and completely inactivated by N-bromosuccinimide(NBS) and slightly decreased by succinic anhydride and N-acetylimidazole. These results indicate that tryptophanyl, lysyl, and tyrosyl residues play an important role in enzyme catalysis. The reaction of alliinase with NBA yielded a characteristic decrease in both the absorbance at 280 nm and the intrinsic fluorescence at 332 nm with increasing reagent concentration of NBS, consistent with the oxidation of tryptophan residues. Kinetic analysis, fluorometric titration of tryptophans and correlation to residual alliinase activity showed that modification of only one residue present on alliinase led to complete inhibition of alliinase activity. To identify this essential tryptophan residue, we employed chemical modification by NBS in the presence and absence of the protecting substrate analogue, S-ethyl-L-cysteine (SEC) and N-terminal sequence analysis of peptide fragment isolated by reverse phase-HPLC. A fragment containing residues 179-188 was isolated. We conclude that Trp182 is essential for alliinase activity.

Development of Surface Cleaning Techniques for Analysis of Electronics Structure in CuInSe2, CuGaSe2 Solar Cell Absorber Layer (태양전지용CuInSe2와 CuGaSe2 흡수층의 전자구조해석을 위한 표면 청정기술 개발)

  • Kim, Kyung-Hwan;Choi, Hyung-Wook;Kong, Sok-Hyun
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.18 no.2
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    • pp.125-129
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    • 2005
  • Two kinds of physical treatments were examined for the analysis both of intrinsic surface and interior nature of CuInS $e_2$[CIS] and CuGaS $e_2$[CGS] films grown in separated systems. For the first method, a selenium protection layer which was immediately deposited after the growth of the CIS was investigated. The Se cap layer protects CISe surface from oxidation and contamination during the transport under ambient atmosphere. The Se cap was removed by thermal annealing at temperature above 15$0^{\circ}C$. After the decapping treatment at 2$25^{\circ}C$ for 60 min, ultraviolet photoemission and inverse photoemission measurements of the CIS film showed that its valence band maximum(VBM) and conduction band minimum (CBM) are located at 0.58 eV below and 0.52 eV above the Fermi level $E_{F}$, respectively. For the second treatment, an Ar ion beam etching was exploited. The etching with ion kinetic energy $E_{k}$ above 500 eV resulted in broadening of photoemission spectra of core signals and occasional development of metallic feature around $E_{F}$. These degradations were successfully suppressed by decreasing $E_{k}$ below 400 eV. CGS films etched with the beam of $E_{k}$ = 400 eV showed a band gap of 1.7 eV where $E_{F}$ was almost centered.st centered.

Formation of DNA-Protein Crosslink at Oxidized Abasic Site Mediated by Human DNA Polymerase Iota and Mitochondrial DNA Polymerase Gamma

  • Son, Mi-Young;Jun, Hyun-Ik;Goo, Sun-Young;Sung, Jung-Suk
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.1-8
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    • 2009
  • Human genomic DNA is continuously attacked by oxygen radicals originated from cellular metabolic processes and numerous environmental carcinogens. 2-deoxyribonolactone (dL) is a major type of oxidized abasic (AP) lesion implicated in DNA strand scission, mutagenesis, and formation of covalent DNA-protein crosslink (DPC) with DNA polymerase (Pol) ${\beta}$. We show here that human DNA polymerase (Pol)${\iota}$ and mitochondrial $Pol{\gamma}$ give rise to stable DNA-protein crosslink (DPC) formation that is specifically mediated by dL lesion. $Pol{\gamma}$ mediates DPC formation at the incised dL residue by its 5'-deoxyribose-5-phosphate (dRP) lyase activity, while $Pol{\gamma}$ cross links with dL thorough its intrinsic dRP lyase and AP lyase activities. Reactivity in forming dL-mediated DPC was significantly higher with $Pol{\gamma}$ than with $Pol{\iota}$. DPC formation by $Pol{\gamma}$, however, can be reduced by an accessory factor of $Pol{\gamma}$ holoenzyme that may attenuate deleterious effects of crosslink adducts on mitochondrial DNA. Comparative kinetic analysis of DPC formation showed that the rate of DPC formation with either $Pol{\iota}$ or $Pol{\gamma}$ was lower than that with $Pol{\beta}$. These results revealed that the activity of catalytic lyase in DNA polymerases determine the efficiency of DPC formation with dL damages. Irreversible crosslink formation of such DNA polymerases by dL lesions may result in a prolonged strand scission and a suicide of DNA repair proteins, both of which could pose a threat to the genetic and structural integrity of DNA.

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Video Event Analysis and Retrieval System for the KFD Web Database System (KFD 웹 데이터베이스 시스템을 위한 동영상 이벤트 분석 및 검색 시스템)

  • Oh, Seung-Geun;Im, Young-Hee;Chung, Yong-Wha;Chang, Jin-Kyung;Park, Dai-Hee
    • The Journal of the Korea Contents Association
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    • v.10 no.11
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    • pp.20-29
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    • 2010
  • The typical Kinetic Family Drawing (KFD) Web database system, a form of prototype system, has been developed, relying on the suggestions from family art therapists, with an aim to handle large amounts of assessment data and to facilitate effective implement of assessment activities. However, unfortunately such a system has an intrinsic problem that it fails to collect clients' behaviors, attitudes, facial expressions, voices, and other critical information observed while they are drawing. Accordingly we propose the ontology based video event analysis and video retrieval system in this paper, in order to enhance the function of a KFD Web database system by using a web camera and drawing tool. More specifically, a newly proposed system is designed to deliver two kinds of services: the client video retrieval service and the sketch video retrieval service, accompanied by a summary report of occurred events and dynamic behaviors relative to each family member object, respectively. The proposed system can support the reinforced KFD assessments by providing quantitative and subjective information on clients' working attitudes and behaviors, and KFD preparation processes.

Efficient Prediction of Broadband Noise of a Centrifugal Fan Using U-FRPM Technique (U-FRPM 기법을 이용한 원심팬 광대역소음의 효율적 예측)

  • Heo, Seung;Cheong, Chulung
    • The Journal of the Acoustical Society of Korea
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    • v.34 no.1
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    • pp.36-45
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    • 2015
  • Recently, a lot of studies have been made about the methods used to generate turbulent velocity fields stochastically in order to effectively predict broadband flow noise. Among them, the FRPM (Fast Random Particle Mesh) method which generates turbulence with specific statistical properties using turbulence kinetic energy and dissipation obtained from the steady solution of the RANS (Reynolds Averaged Navier-Stokes) equations has been successfully applied. However, the FRPM method cannot be applied to the flow noise problems involving intrinsic unsteady characteristics such as centrifugal fan. In this paper, to effectively predict the broadband noise generated by centrifugal fan, U-FRPM (unsteady FRPM) method is developed by extending the FRPM method to be combined with the unsteady numerical solutions of the unsteady RANS equations to generate the turbulence considered as broadband noise sources. Firstly, an unsteady flow field is obtained from the unsteady RANS equations through CFD (Computational Fluid Dynamics). Then, noise sources are generated using the U-FRPM method combined with acoustic analogy. Finally, the linear propagation model which is realized through BEM (Boundary Element Method) is combined with the generated sources to predict broadband noise at the listeners' position. The proposed technique is validated to compare its prediction result with the measured data.

Sulfate reduction and sulfur isotopic fractionation in marine sediments (해양퇴적물내에서의 황산염 환원과 황의 안정동위원소 분화)

  • 한명우
    • Journal of Environmental Science International
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    • v.2 no.1
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    • pp.43-49
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    • 1993
  • Concentrations of sulfate and 6-values of sulfate, $({\delta}^{34}SO_4_){pw}$, dissolved In pore waters were measured from the sediment cores of the two different marine environments : deep northeast Pacific (57-1) and coastal Kyunggi Bay of Yellow Sea (57-2) . Sulfate concentration in pore waters decreases with depth at both cores, reflecting sulfate reduction in the sediment columns. However, much higher gradient of pore water sulfate at 57-2 than 57-1 indicates more rapid sulfate reduction at 57-2, because of high sedimentation rate at the coastal area compared to the deep-sea. The measured 6-values, $({\delta}^{34}SO_4_){pw}$, follow extremely well the predicted trend of the Rayleigh fractionation equation. The range of 26.756 to 61.35% at the coastal core 57-2 is not so great as that of 32.4$\textperthousand$ to 97.8$\textperthousand$ at the deep-sea core 57-1. Despite greater graclient of pore water sulfate at 57-2, the 6-values become lower than those of the deep- sea core 57-1. This inverse relation between the 6-values and the gradients of pore water sulfate could be explained by the combination of the two subsequent factors : the kinetic effect by which the residual pore water sulfate becomes progressively enriched with respect to the heavy isotope of $^{34}S$ as sulfate reduction proceeds, and the intrinsic formulation effect of the Rayleigh fractionation equation in which the greater becomes the fractionation factor, the more diminished values of $({\delta}^{34}SO_4_){pw}$ are predicted.

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