• Title/Summary/Keyword: binding constant

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Cloning, Over-expression, and Characterization of YjgA, a Novel ppGpp-binding Protein

  • Gnanasekaran, Gopalsamy;Pan, SangO;Jung, Wontae;Jeong, Kwangjoon;Jeong, Jae-Ho;Rhee, Joon Haeng;Choy, Hyon E.;Jung, Che-Hun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2419-2424
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    • 2013
  • Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

Fluorescence Sensing Properties of 2-(2'-Hydroxyphenyl)quinoline and Derivatives

  • Helal, Aasif;Lee, Sang-Hoon;Ren, Wen Xiu;Cho, Chan-Sik;Kim, Hong-Seok
    • Bulletin of the Korean Chemical Society
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    • v.32 no.5
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    • pp.1599-1603
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    • 2011
  • Novel chemosensors based on 2-(2'-hydroxyphenyl)quinoline were prepared and evaluated for sensing metal cations. The photophysical properties of chemosensors 1-3 were examined and their ion-selectivity was evaluated by measuring their fluorescent emission responses to alkali, alkaline earth, and transition metal ions. Chemosensors 1, 2 and 3 show ratiometric and enhanced fluorescence changes with transitional metals that are efficient fluorescence quenchers, especially 3 has a high binding constant with $Hg^{+2}$ in $CH_3CN$.

A Theoretical Study of the Formation of Benzene Excimer: Effects of Geometry Relaxation and Spin-state Dependence

  • Kim, Dongwook
    • Bulletin of the Korean Chemical Society
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    • v.35 no.9
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    • pp.2738-2742
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    • 2014
  • Geometry relaxation effects on the formation of benzene excimer were investigated by means of ab initio calculation at SOS-CIS($D_0$)/aug-cc-pVDZ level. In the case of T-shaped dimer configuration, intermolecular interactions in the excited states are found to be nearly the same as those in the ground state and structural deformations are limited within a single molecule; the geometry relaxation effects are then negligible and singlet-triplet energy gap remains constant. As for face-to-face eclipsed dimer, on the other hand, both molecules undergo structural change. As a result, intermolecular interactions in the excited states are significantly different than those in the ground state. Although the intermolecular distances obtained from potential energy curve calculation with frozen molecular structures are in qualitative agreement, the excited-state binding energies are notably overestimated with respect to those at optimized structures. In particular, the effects are calculated to be larger in $T_1$ state and hence singlet-triplet energy gap, which reduces markedly in this configuration, is underestimated without relaxation.

Synthesis of Facial Amphiphile 3,7-Diamino-5α-cholestane Derivatives as a Molecular Receptor

  • Ahmad, Md. Wasi;Jung, Young-Mee;Khan, Sharaf Nawaz;Kim, Hong-Seok
    • Bulletin of the Korean Chemical Society
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    • v.30 no.9
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    • pp.2101-2106
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    • 2009
  • A series of facial amphiphiles 3,7-diaminocholestane were synthesized from 3,7-diketocholestane via 2 sequential reductive aminations and anion recognition was evaluated with acetate, chloride, bromide, fluoride and phosphate anions. The stereo-selective reductive amination protocol was utilized to synthesized facial amphiphiles afforded receptors in high yields. The molecular receptor 2 showed the highest binding constant with acetate in a 1:1 ratio.

Diethylpyrocarbonate Inactivation of Aspartase from Hafnia Alvei

  • Shim, Jae-Hee;Kim, Hyo-Joon;Yoon, Moon-Young
    • BMB Reports
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    • v.32 no.4
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    • pp.326-330
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    • 1999
  • An aspartase purified from Hafnia alvei was inactivated by diethylpyrocarbonate (DEP) in a pseudo-first-order inactivation. The first-order plot was biphasic. The inactivation process was not saturable and the second order rate constant was $1.3\;M^{-1}s^{-1}$. The inactivated aspartase was reactivated with NH₂OH. The difference absorption spectrum of DEP-inactivated vs native enzyme preparations revealed a marked peak around 242 nm. The pH dependence of the inactivation rate suggests that an amino acid residue having a pK value of 7.2 was involved in the inactivation. L-aspartate, fumarate (substrates), and chloride ion (inhibitor) protected the enzyme against inactivation, indicating that histidine residues for the enzyme activity are located at the active site of this aspartase. Inspection of the presence and absence of $Cl^-$ ion demonstrated that the number of essential histidine residues is less than two. Thus, one or two histidines are in or near the aspartate binding site and participate in an essential step of the catalytic reaction.

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Synthesis and Photoaffinity Labeling of 3'(2')-O-(p-azidobenzoyl) ATP

  • Shin, Seung-Jin;Lee, Woo-Kyoung;Park, Jong-Sang
    • BMB Reports
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    • v.30 no.3
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    • pp.211-215
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    • 1997
  • A photoactive analog of ATP, 3'(2')-O-(p-azidobenzoyl)-adenosine 5-triphosphate (AB-ATP) was synthesized by chemically coupling N-hydroxysuccinimidyl-4-azidobenzoate (NHS-AB) and ATP. The utility of AB-ATP as an effective active-site-directed photoprobe was demonstrated using catalytic subunit of protein kinase A as a model enzyme. Photoincorporation of AB-ATP was saturated with apparent dissociation constant of $30{\mu}m$ and protected completely by $100{\mu}m$ of ATP. When the enzyme was covalently modified by photolysis in the presence of saturating amounts of photoprobe, about 60% inhibition of enzyme activity was observed. These results demonstrate that AB-ATP has potential application as a probe to characterize ATP-binding proteins including protein kinases.

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Tyrosinase Inhibitory Activities of Safrole from Myristica fragrans Houtt.

  • Cho, Soo Jeong;Kwon, Hyun Sook
    • Journal of Applied Biological Chemistry
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    • v.58 no.4
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    • pp.295-301
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    • 2015
  • Five phenylpropanoids (1-5), a benzofuran neolignan (6), two 8-O-4'-neolignans (7-8), and five tetrahydrofuran lignans (9-13) were isolated from a methanol extract of Myristica fragrans seeds. The structures of 1-13 were determined by $^1H$- and $^{13}C$-NMR spectroscopic data analyses and a comparison with the literature data. Compound 3 was isolated for the first time from this plant. All the isolated compounds were evaluated for their inhibitory activity against tyrosinase. Among them, safrole (1) showed significant inhibitions against both the monophenolase ($IC_{50}=32.11{\mu}M$) and diphenolase ($IC_{50}=27.32{\mu}M$) activities of tyrosinase. The kinetic analysis shows that safrole (1) is competitive inhibitors for both monophenolase and diphenolase. The apparent inhibition constant ($K_i$) for safrole (1) binding with free enzyme was determined to be 16.05 and $13.66{\mu}M$ for monophenolase and diphenolase, respectively.

Simple Pyrene Derivatives as Fluorescence Sensors for TNT and RDX in Micelles

  • Hong, Jung-Ho;Choi, Jung-Hwa;Cho, Dong-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.35 no.11
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    • pp.3158-3162
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    • 2014
  • Various pyrene derivatives were synthesized and systematically examined in micelles. Synthesized mono and bispyrene derivatives were tested in micelles so that they displayed a strong excimer band and the excimer band was quenched in the presence of TNT and RDX. In the optimized condition, the binding constant for TNT of a simple dipyrene derivative 4 was increased up to $1.0{\times}10^6M^{-1}$ in cetyl trimethylammonium bromide (CTAB) micelles, which allowed for the detection of 2 ppb of TNT and 334 ppb of RDX by fluorescence titrations.

Characteristics of Phosphors for PDP with Frit Contents (Frit 첨가량에 따른 PDP용 형광체의 특성 연구)

  • Jung, Ah-Reum;Kim, Hyeong-Jun;Choi, Sung-Churl
    • Journal of the Korean Ceramic Society
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    • v.47 no.2
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    • pp.146-150
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    • 2010
  • Because the plasma display panel has used red, green and blue(RGB) phosphors, it has suffer from two intrinsic problems; 1) the cell defect due to the lack of binding force between phosphor particles and 2) mis-discharge because of difference of electrical characteristics among RGB phosphors. In order to control the mechanical and electrical properties of RGB phosphors, frit with $ZnOB_2O_3-SiO_2-Al_2O_3$ system was added to RGB phosphor as sintering additive. The mechanical properties were increased by the amount of frit. The amount of frit under 5 wt% rarely affected dielectric constant. However, there was the limit of amount because of decreasing optical properties seriously; over 3 wt% in red, over 10 wt% in green and blue.

Interband optical properties in wide band gap group-III nitride quantum dots

  • Bala, K. Jaya;Peter, A. John
    • Advances in nano research
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    • v.3 no.1
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    • pp.13-27
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    • 2015
  • Size dependent emission properties and the interband optical transition energies in group-III nitride based quantum dots are investigated taking into account the geometrical confinement. Exciton binding energy and the optical transition energy in $Ga_{0.9}In_{0.1}N$/GaN and $Al_{0.395}In_{0.605}N$/AlN quantum dots are studied. The largest intersubband transition energies of electron and heavy hole with the consideration of geometrical confinement are brought out. The interband optical transition energies in the quantum dots are studied. The exciton oscillator strength as a function of dot radius in the quantum dots is computed. The interband optical absorption coefficients in GaInN/GaN and AlInN/AlN quantum dots, for the constant radius, are investigated. The result shows that the largest intersubband energy of 41% (10%) enhancement has been observed when the size of the dot radius is reduced from $50{\AA}$ to $25{\AA}$ of $Ga_{0.9}In_{0.1}N$/GaN ($Al_{0.395}In_{0.605}N$/AlN) quantum dot.