• Title/Summary/Keyword: metal binding

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Mutational Analysis of an Essential RNA Stem-loop Structure in a Minimal RNA Substrate Specifically Cleaved by Leishmania RNA Virus 1-4 (LRV1-4) Capsid Endoribonuclease

  • Ro, Youngtae;Patterson, Jean L.
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.239-247
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    • 2003
  • The LRV1-4 capsid protein possesses an endoribonuclease activity that is responsible for the single site-specific cleavage in the 5' untranslated region (UTR) of its own viral RNA genome and the formation of a conserved stem-loop structure (stem-loop IV) in the UTR is essential for the accurate RNA cleavage by the capsid protein. To delineate the nucleotide sequences, which are essential for the correct formation of the stem-loop structure for the accurate RNA cleavage by the viral capsid protein, a wildtype minimal RNA transcript (RNA 5' 249-342) and several synthetic RNA transcripts encoding point-mutations in the stem-loop region were generated in an in vitro transcription system, and used as substrates for the RNA cleavage assay and RNase mapping studies. When the RNA 5' 249-342 transcript was subjected to RNase T1 and A mapping studies, the results showed that the predicted RNA secondary structure in the stem-loop region using FOLD analysis only existed in the presence of Mg$\^$2+/ ions, suggesting that the metal ion stabilizes the stem-loop structure of the substrate RNA in solution. When point-mutated RNA substrates were used in the RNA cleavage assay and RNase T1 mapping study, the specific nucleotide sequences in the stem-loop region were not required for the accurate RNA cleavage by the viral capsid protein, but the formation of a stem-loop like structure in a region (nucleotides from 267 to 287) stabilized by Mg$\^$2+/ ions was critical for the accurate RNA cleavage. The RNase T1 mapping and EMSA studies revealed that the Ca$\^$2+/ and Mn$\^$2+/ ions, among the reagents tested, could change the mobility of the substrate RNA 5' 249-342 on a gel similarly to that of Mg$\^$2+/ ions, but only Ca$\^$2+/ ions identically showed the stabilizing effect of Mg$\^$2+/ ions on the stem-loop structure, suggesting that binding of the metal ions (Mg$\^$2+/ or Ca$\^$2+/) onto the RNA substrate in solution causes change and stabilization of the RNA stem-loop structure, and only the substrate RNA with a rigid stem-loop structure in the essential region can be accurately cleaved by the LRV1-4 viral capsid protein.

Surface Modification Using Spiropyran-Derivative and Its Analysis of Surface Potential Induced by UV (스파이로파이란에 의한 표면 개질 및 자외선에 의해 유도된 표면 전위에 대한 분석)

  • Lee, Bong-Soo;Han, Dong-Keun;Son, Tae-Il;Jung, Young-Hwan
    • Journal of the Korean Chemical Society
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    • v.55 no.3
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    • pp.478-485
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    • 2011
  • Merocyanine derivatives transformed from spiropyran-containing compounds by irradiating the light of ultraviolet (UV) include zwitterion of phenolate anion and amine cation. Complexation of this phenolate anion on merocyaninemodified surface and Ni ion among metal ions led to a change of surface charge and it was measured with kelvin prove force microscopy (KFM). We found that the resultant surface potential decreased linearly as UV-exposed time increased, and finally were saturated. Also it was analyzed through XPS the immobilized amount of Ni ions was increased according to increase of UV-exposed time. It is considered that these properties could be applied for detection and a quantitative control of different metal ions. Further research is to aim construct specific scaffold/matrix which enable high selective, high sensitive and, especially, a quantitative immobilization of metal ions-binding biomaterials such as proteins and cells.

Variations in Heavy Metal Analytical Results and Leaching Characteristics of Coal Ash Recycled Concretes according to Sample Crushing Methods (분쇄방법에 따른 석탄재 재활용 콘크리트의 중금속 분석결과 및 용출특성 변화)

  • Lee, Jin Won;Choi, Seung-Hyun;Kim, Kangjoo;Moon, Bo-Kyung
    • Economic and Environmental Geology
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    • v.51 no.5
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    • pp.429-438
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    • 2018
  • Since concrete is a hardened aggregates of various materials, it needs to be crushed for chemical analyses. However, the effect of sample crushing on the analytical results has not been precisely assessed till today. In this study, we prepared concrete test pieces using Portand cements and fly ashes as binding materials, and ponded ashes and sands as aggregates and analyzed the heavy metals of the test pieces using Standards for Fair Testing of Soil Contamination (SFTSC) and Wastes (SFTW). For this, each test piece was partially crushed at first and sieved for separation of grains of <0.15 mm, 0.15-0.5 mm, and 0.5-5 mm from the same crushed samples (Crushing Method I). Results of those samples using SFTSC showed a clear trend that analyzed heavy metal concentrations are higher in the finer fractions. Particularly, fractions with <0.15 mm indicated much higher concentrations than the theoretical ones, which were calculated based on the concentrations of individual materials and their mixing fractions. In contrast, the analytical results were generally comparable with the theoretical ones when the test pieces were totally pulverized such that all the crushed grains were <0.15 mm in size (Crushing Method II). These results are associated with the fact that cement materials and fly ashes, which are high in heavy metals relative to other materials, are enriched in the fine fractions. The analytical results using the SFTW derived very low concentrations in most of parameters and did not indicate the dependence of concentrations on the crushing methods due to using distilled water as leaching agent.

Characteristics of Growth and Metal Removal in Recombinant Saccharomyces cerevisiae harboring a Metallothionein Gene (Metallothionein 유전자가 도입된 재조합 Saccharomyces cerevisiae의 생육과 금속제거에 대한 특성)

  • 정동환;김대옥서진호
    • KSBB Journal
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    • v.10 no.5
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    • pp.475-481
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    • 1995
  • The effect of metallothionein expression on the metal resistance and removal by recombinant Saccharomyces cerevisiae containing the plasmid pJW9 was investigated. The recombinant strain S. cerevisiae BZ-pJ was constructed by transforming the host strain S. cerevisiae BZ3l-1-7Ba with the gene coding for a metal-binding protein, metallothionein. Introduction of the MT gene yielded an increase in the minimum inhibitory concentration (MIC) of copper more than three times compared with the host strain. The minimum inhibitory concentrations of $Cr^{2+}, Znr^{2+} and Pb^{2+}, $ were not different for the two strains. The recombinant yeast grown in a medium containing 8mM CuSO4 was able to remove copper with a capacity of 18.9mg $Cu^{2+}$/g dry cell. In a mixture of copper and zinc, the presence of copper relieved the toxic effects caused by zinc, resulting in an enhancement of the final cell density and the specific growth rate of the recombinant yeast. The capability to remove copper by the recombinant yeast was linearly proportional to the copper concentrations in the medium. The efficiency of copper removal was rather constant regardless of the initial copper concentrations. The specific removal of zinc was dependent on the zinc concentrations in media, though, and such dependence was not so pronounced as the concentration of copper.

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Adherence of Salivary Proteins to Various Orthodontic Brackets (다양한 교정용 브라켓 표면에 부착하는 타액단백질에 관한 연구)

  • Ahn, Sug-Joon;Ihm, Jong-An;Nahm, Dong-Seok
    • The korean journal of orthodontics
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    • v.32 no.6 s.95
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    • pp.443-453
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    • 2002
  • The principal aims of this study were to identify the composition of salivary pellicles formed on various orthodontic brackets and to obtain a detailed information about the protein adsorption profiles from whole saliva and two major glandular salivas. Four different types of orthodontic brackets were used. All were upper bicuspid brackets with a $022{\times}028$ slot Roth prescription; stainless steel metal, monocrystalline sapphire, polycrystalline alumina, and plastic brackets. Bracket pelicles were formed by the incubation of orthodontic brackets with whole saliva, submandibular-sublingual saliva, and parotid saliva for 2 hours. The bracket pellicles were extracted and confirmed by employing sodium dodecyl sulfatepolyacrylamide gel electrophoresis, Western transfer methods, and immunodetection. The results showed that low-molecular weight salivary mucin, ${\alpha}-amylase$, secretory IgA (sIgA), acidic proline-rich proteins, and cystatins were attached to all of these brackets regardless of the bracket types. High-molecular weight mucin, which promotes the adhesion of Streptococcus mutans, did not adhere to uy orthodontic brackets. Though the same components were detected in all bracket pellicles, however, the gel profiles showed qualitatively and quantitatively different pellicles, according to the origins of saliva and the bracket types. In particular, the binding of sIgA was more prominent in the pellicles from parotid saliva and the binding of cystatins was prominent in the pellicles from the form plastic brackets. This study indicates that numerous salivary proteins adhere to the orthodontic brackets and these salivary proteins adhere selectively according to bracket types and the types of the saliva.

Enhanced Catalytic Activity of Cu/Zn Catalyst by Ce Addition for Low Temperature Water Gas Shift Reaction (Ce 첨가에 따른 저온수성가스전이반응용 Cu/Zn 촉매의 활성 연구)

  • Byun, Chang Ki;Im, Hyo Bin;Park, Jihye;Baek, Jeonghun;Jeong, Jeongmin;Yoon, Wang Ria;Yi, Kwang Bok
    • Clean Technology
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    • v.21 no.3
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    • pp.200-206
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    • 2015
  • In order to investigate the effect of cerium oxide addition, Cu-ZnO-CeO2 catalysts were prepared using co-precipitation method for water gas shift (WGS) reaction. A series of Cu-ZnO-CeO2 catalyst with fixed Cu Content (50 wt%, calculated as CuO) and a given ceria content (e.g., 0, 5, 10, 20, 30, 40 wt%, calculated as CeO2) were tested for catalytic activity at a GHSV of 95,541 h-1, and a temperature range of 200 to 400 ℃. Cu-ZnO-CeO2 catalysts were characterized by using BET, SEM, XRD, H2-TPR, and XPS analysis. Varying composition of Cu-ZnO-CeO2 catlysts led the difference characteristics such as Cu dispersion, and binding energy. The optimum 10 wt% doping of cerium facilitated catalyst reduction at lower temperature and improved the catalyst performance greatly in terms of CO conversion. Cerium oxide added catalyst showed enhanced activities at higher temperature when it compared with the catalyst without cerium oxide. Consequently, ceria addition of optimal composition leads to enhanced catalytic activity which is attributed to enhanced Cu dispersion, lower binding energy, and hindered Cu metal agglomeration.

Soluble Prokaryotic Expression and Purification of Bioactive Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand

  • Do, Bich Hang;Nguyen, Minh Tan;Song, Jung-A;Park, Sangsu;Yoo, Jiwon;Jang, Jaepyeong;Lee, Sunju;So, Seoungjun;Yoon, Yejin;Kim, Inki;Lee, Kyungjin;Jang, Yeon Jin;Choe, Han
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2156-2164
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    • 2017
  • Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

Photodecomposition of Concentrated Ammonia over Nanometer-sized TiO2, V-TiO2, and Pt/V-TiO2 Photocatalysts

  • Choi, Hyung-Joo;Kim, Jun-Sik;Kang, Mi-Sook
    • Bulletin of the Korean Chemical Society
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    • v.28 no.4
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    • pp.581-588
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    • 2007
  • To enhance the photodecomposition of concentrated ammonia into N2, Pt/V-TiO2 photocatalysts were prepared using solvothermal and impregnation methods. Nanometer-sized particles of 0.1, 0.5 and 1.0 mol% V-TiO2 were prepared solvothermally, and then impregnated with 1.0 wt% Pt. The X-ray diffraction (XRD) peaks assigned to V2O5 at 30.20 (010) and Pt metal at 39.80 (111) and 46.20 (200) were seen in the 1.0 wt% Pt/ 10.0 mol% V-TiO2. The particle size increased in the order: pure TiO2, V-TiO2 and Pt/V-TiO2 after thermal treatment at 500 °C, while their surface areas were in the reverse order. On X-ray photoelectron spectroscopy (XPS), the bands assigned to the Ti2p3/2 and Ti2p1/2 of Ti4+-O were seen in all the photocatalysts, and the binding energies increased in the order: TiO2 < Pt/V-TiO2 < V-TiO2. The XPS bands assigned to the V2p3/2 (517.85, 519.35, and 520.55 eV) and V2p1/2 (524.90 eV) in the V3+, V4+ and V5+ oxides appeared over V-TiO2, respectively, while the band shifted to a lower binding energy with Pt impregnation. The Pt components of Pt/ V-TiO2 were identified at 71.60, 73.80, 75.00 and 76.90 eV, which were assigned to metallic Pt 4f7/2, PtO 4f7/2, PtO2 4f7/2, and PtO 4f5/2, respectively. The UV-visible absorption band shifted closer towards the visible region of the spectrum in V-TiO2 than in pure TiO2 and; surprisingly, the Pt/V-TiO2 absorbed at all wavelengths from 200 to 800 nm. The addition of vanadium generated a new acid site in the framework of TiO2, and the medium acidic site increased with Pt impregnation. The NH3 decomposition increased with the amount of vanadium compared to pure TiO2, and was enhanced with Pt impregnation. NH3 decomposition of 100% was attained over 1.0 wt% Pt/1.0 mol% V-TiO2 after 80 min under illumination with 365 nm light, although about 10% of the ammonia was converted into undesirable NO2 and NO. Various intermediates, such as NO2, -NH2, -NH and NO, were also identified in the Fourier transform infrared (FT-IR) spectra. From the gas chromatography (GC), FT-IR and GC/mass spectroscopy (GC/MS) analyses, partially oxidized NO and NO2 were found to predominate over V-TiO2 and pure TiO2, respectively, while both molecules were reduced over Pt/V-TiO2.

Pd/Pd3Fe Alloy Catalyst for Enhancing Hydrogen Production Rate from Formic Acid Decomposition: Density Functional Theory Study (개미산 분해 반응에서 수소 생산성 증대를 위한 Pd/Pd3Fe 합금 촉매: 범밀도 함수 이론 연구)

  • Cho, Jinwon;Han, Jonghee;Yoon, Sung Pil;Nam, Suk Woo;Ham, Hyung Chul
    • Korean Chemical Engineering Research
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    • v.55 no.2
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    • pp.270-274
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    • 2017
  • Formic acid has been known as one of key sources of hydrogen. Among various monometallic catalysts, hydrogen can be efficiently produced on Pd catalyst. However, the catalytic activity of Pd is gradually reduced by the blocking of active sites by CO, which is formed from the unwanted indirect oxidation of formic acid. One of promising solutions to overcome such issue is the design of alloy catalyst by adding other metal into Pd since alloying effect (such as ligand and strain effect) can increase the chance to mitigate CO poisoning issue. In this study, we have investigated formic acid deposition on the bimetallic $Pd/Pd_3Fe$ core-shell nanocatalyst using DFT (density functional theory) calculation. In comparison to Pd catalyst, the activation energy of formic acid dehydrogenation is greatly reduced on $Pd/Pd_3Fe$ catalyst. In order to understand the importance of alloying effects in catalysis, we decoupled the strain effect from ligand effect. We found that both strain effect and ligand effect reduced the binding energy of HCOO by 0.03 eV and 0.29 eV, respectively, compared to the pure Pd case. Our DFT analysis of electronic structure suggested that such decrease of HCOO binding energy is related to the dramatic reduction of density of state near the fermi level.

Synthesis of Diketo Copper(II) Complex and Its Binding toward Calf Thymus DNA (CTDNA) (이케토 구리(II) 착물의 합성 및 송아지 Thymus DNA(CTDNA)와의 상호작용)

  • Tak, Aijaz Ahmad;Arjmand, Farukh
    • Journal of the Korean Chemical Society
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    • v.55 no.2
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    • pp.177-182
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    • 2011
  • A diketo-type ligand was synthesized by the Knoevenagel condensation reaction of thiophene-2-aldehyde with acetylacetone, subsequently its transition metal complexes with Cu(II), Ni(II), and Co(II) chlorides were also prepared. All the complexes were characterized by various physico-chemical methods. The molar conductivity data reveals ionic nature for the complexes. The electronic spectrum and the EPR values suggest square planar geometry for the Cu(II) ion. Interaction of the Cu(II) complex with CTDNA (calf thymus DNA) was studied by absorption spectral method and cyclic voltammetry. The $k_{obs}$ values versus [DNA] gave a linear plot suggesting psuedo-first order reaction kinetics. The cyclic voltammogram of the Cu(II) complex reveals a quasi-reversible wave attributed to Cu(II)/Cu(I) redox couple for one electron transfer with $E_{1/2}$ values -0.240 V and -0.194 V. respectively. On addition of CTDNA, there is a shift in the $E_{1/2}$ values 168 mV and 18 mV respectively and decrease in Ep values. The shift in $E_{1/2}$ values in the presence of CTDNA suggests strong binding of Cu(II) complex to the CTDNA.