• Title/Summary/Keyword: Quencher

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Conformation of Single Stranded Poly(dA) and Its Interaction with 4',6-Diamidino-2-phenylindole

  • 정갑선;김명순;이길준;조태섭;Kim, Seog K.;이세윤
    • Bulletin of the Korean Chemical Society
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    • v.18 no.5
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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).

pH and Micellar Effects on the Quenching of Tris(2,2'-bipyridine)Ruthenium(Ⅱ) Luminescence by 1-Alkyl-4,4'-bipyridinium : Evidence of Deep Enbedment of the Quencher Cations in Sodium Dodecyl Sulfate Micelle

  • 박준우;김유나
    • Bulletin of the Korean Chemical Society
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    • v.18 no.2
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    • pp.185-189
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    • 1997
  • The effect of pH and sodium dodecyl sulfate (SDS) micelle on the quenching of Ru(bpy)32+ luminescence by N-alkyl-4,4'-bipyridinium ions (RBPY+: R=methyl, octyl, dodecyl, benzyl) were investigated. In the absence of SDS, the quenching rate at pH 2 is similar to that of the corresponding methylalkyl viologens and much greater than that in pH 8 solution due to greater reducibility of the protonated form of RBPY+ (HRBPY2+). The quenching rate at pH 2 is strongly enhanced by the presence of SDS, while that at basic pH is greatly retarded. These observations are explained by deep embedment of RBPY+ into the hydrophobic hydrocarbon region of the micelle, whereas Ru(bpy)32+ and HRBPY2+ locate in the Stern layer of the micelle.

Oligomerization State of the Plasma Membrane Proteolipid Apoprotein Purified from the Bovine Kidney, Probed by the Fluorescence Polarization

  • Chae, Quae;Nam, Sang-Rye
    • Bulletin of the Korean Chemical Society
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    • v.9 no.4
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    • pp.202-206
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    • 1988
  • In order to investigate the oligomerization state of the plasma membrane proteolipid apoprotein purified from the bovine kidney, fluorescence polarization experiment was carried out in the two different solvent systems, i.e., water and organic solvent(chloroform-methanol). The molecular volumes of the proteins estimated from the Perrin equation, were to be 45,258$A^3$ and 17,608$A^3$ in water and organic solvent, respectively. These values indicate that a trimerization is possibly occurring in the aqueous environment. As an auxiliary experiment for the calculation of the molecular volume using Perrin equation, fluorescence quenching constants ($K_q$) with the quencher acrylamide and fluorescence lifetimes (${\tau}_F$) of the intrinsic fluorophore tryptophan residue were estimated in the two different solvent systems. $K_q$ in water was 18.21$M^{-1}$ and it was 46.24$M^{-1}$ in organic solvent. Fluorescence lifetimes of tryptophan residue were calculated to be 2.80 nsec. in water and 3.81 nsec. in organic solvent, respectively.

Ionic Strength Dependent Binding Mode of 9-Aminoacridine to DNA

  • 김혜경;조태섭;Kim, Seog K.
    • Bulletin of the Korean Chemical Society
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    • v.17 no.4
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    • pp.358-362
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    • 1996
  • The ionic strength dependent binding mode of 9-aminoacridine (9AA), a well-known DNA intercalator, to DNA is studied by flow linear dichroism, circular dichroism, fluorescence techniques and equilibrium dialysis. The DNA-bound 9AA exhibits spectral properties corresponding to the intercalative binding mode disregarding the salt concentrations; the angle between the long-axis transition moment of the 9AA molecule and DNA helix axis is calculated to be about 65°, indicating a significant deviation from the classical intercalation. At low salt concentrations, however, upwards bending curve in Stern-Volmer plot is observed (where 9AA is a fluorophore and DNA a quencher), indicating the coexistence of both static and dynamic quenching mechanisms or the existence of an additional binding site.

Sequence Dependent Binding Modes of the ΔΔ- and ΛΛ-binuclear Ru(II) Complexes to poly[d(G-C)2] and poly[d(A-T)2]

  • Chitrapriya, Nataraj;Kim, Raeyeong;Jang, Yoon Jung;Cho, Dae Won;Han, Sung Wook;Kim, Seog K.
    • Bulletin of the Korean Chemical Society
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    • v.34 no.7
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    • pp.2117-2124
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    • 2013
  • The binding properties and sequence selectivities of ${\Delta}{\Delta}$- and ${\Lambda}{\Lambda}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ (bip = 4,4'-biphenylene (imidazo [4,4-f][1,10]phenanthroline) complexes with $poly[d(A-T)_2]$ and $poly[d(G-C)_2]$ were investigated using conventional spectroscopic methods. When bound to $poly[d(A-T)_2]$, a large positive circular dichroism (CD) spectrum was induced in absorption region of the bridging moiety for both the ${\Delta}{\Delta}$- and ${\Lambda}{\Lambda}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ complexes, which suggested that the bridging moiety sits in the minor groove of the polynucleotide. As luminescence intensity increased, decay times became longer and complexes were well-protected from the negatively charged iodide quencher compared to that in the absence of $poly[d(A-T)_2]$. These luminescence measurements indicated that Ru(II) enantiomers were in a less polar environment compared to that in water and supported by minor groove binding. An angle of $45^{\circ}$ between the molecular plane of the bridging moiety of the ${\Delta}{\Delta}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ complex and the local DNA helix axis calculated from reduced linear dichroism ($LD^r$) spectrum further supported the minor groove binding mode. In the case of ${\Lambda}{\Lambda}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ complex, this angle was $55^{\circ}$, suggesting a tilt of DNA stem near the binding site and bridging moiety sit in the minor groove of the $poly[d(A-T)_2]$. In contrast, neither ${\Delta}{\Delta}$-nor ${\Lambda}{\Lambda}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ complex produced significant CD or $LD^r$ signal in the absorption region of the bridging moiety. Luminescence measurements revealed that both the ${\Delta}{\Delta}$- and ${\Lambda}{\Lambda}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ complexes were partially accessible to the $I^-$ quencher. Furthermore, decay times became shorter when bis-Ru(II) complexes bound to $poly[d(G-C)_2]$. These observations suggest that both the ${\Delta}{\Delta}$- and ${\Lambda}{\Lambda}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ complexes bind at the surface of $poly[d(G-C)_2]$, probably electrostatically to phosphate group. The results indicate that ${\Delta}{\Delta}$- and ${\Lambda}{\Lambda}-[{\mu}-Ru_2(phen)_4(bip)]^{4+}$ are able to discriminate between AT and GC base pairs.

Fabrication of Hydrogel and Gas Permeable Membranes for FET Type Dissolved $CO_{2}$ Sensor by Photolithographic Method (사진식각법을 이용한 FET형 용존 $CO_{2}$ 센서의 수화젤막 및 가스 투과막 제작)

  • Park, Lee-Soon;Kim, Sang-Tae;Koh, Kwang-Nak
    • Journal of Sensor Science and Technology
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    • v.6 no.3
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    • pp.207-213
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    • 1997
  • A field effect transistor(FET) type dissolved carbon dioxide($pCO_{2}$) sensor with a double layer structure of hydrogel membrane and $CO_{2}$ gas permeable membrane was fabricated by utilizing a $H^{+}$ ion selective field effect transistor(pH-ISFET) with Ag/AgCl reference electrode as a base chip. Formation of hydrogel membrane with photo-crosslinkable PVA-SbQ or PVP-PVAc/photosensitizer system was not suitable with the photolithographic process. Furthermore, hydrogel membrane on pH-ISFET base chip could be fabricated by photolithographic method with the aid of N,N,N',N'-tetramethyl othylenediarnine(TED) as $O_{2}$ quencher without using polyester film as a $O_{2}$ blanket during UV irradiation process. Photosensitive urethane acrylate type oligomer was used as gas permeable membrane on top of hydrogel layer. The FET type $pCO_{2}$ sensor fabricated by photolithographic method showed good linearity (linear calibration curve) in the range of $10^{-3}{\sim}10^{0}\;mol/{\ell}$ of dissolved $CO_{2}$ in aqueous solution with high sensitivity.

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Research Method of Fatty Acids Transfer between Phospholipid Model Membranes (인지질 모델막에서의 지방산 이동에 관한 연구 방법)

  • 임병순;김혜경;김을상
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.4
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    • pp.743-750
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    • 1997
  • Direct measurement of the kinetics of free fatty acid transfer between phospholipid model membrane is technically limited by the rapid nature of the transfer process. Separation of membrane-bound fatty acid by centrifugation has shown that although the equilibrium distribution of free fatty acid is determined by this method, fatty acid transfer occurs too rapidly for accurate kinetic measurements. Recently fluorescence resonance energy transfer(FRET) assay has been developed to examine transfer of fatty acids between membranes. Donor membranes which has fluorescent fatty acid, anthroyloxy fatty acid(AOFA), is mixed with acceptor membranes which has non-interchangeable fluorescent quencher, nitrobenzo-xadiazol(NBD), using stopped flow apparatus. As the fluorescent fatty acids transfer from donor membrane to acceptor membrane, fluorescence intensity would be decreased and the rate and degree of fatty acid transfer can be analyzed. Fatty acid transfer between micelles is more complicated because of bile salt. Therefore in experiments with micelles, fluorescence self quenching assay is used. At high concentrations, a fluorophore tends to quench its own fluorescence causing a reduction in fluorescence intensity. Donor micelles contained self quenching concentrations of fluorophore and acceptor micelles had no fluorophore. Upon mixing of donor and acceptor micelles, the rate of transfer of the fluorophore from the donor to the acceptor was measured by monitoring the release in self quenching when its concentration in donor decreased over time.

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Spectroscopic Studies on the Mechanism of Interaction of Vitamin $B_{12}$ with Bovine Serum Albumin

  • Kamat, B.P.;Seetharamappa, J.
    • Journal of Photoscience
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    • v.11 no.1
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    • pp.29-33
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    • 2004
  • The mechanism of interaction of cyanocobalamin (CB) with bovine serum albumin (BSA) has been investigated by spectrofluorometric and circular dichroism methods. Association constant for the CB-BSA system showed that the interaction is non-covalent in nature. Binding studies in the presence of an hydrophobic probe, 8-anilino-l-naphthalene sulphonic acid, sodium salt (ANS) showed that there is hydrophobic interaction between CB and ANS and they do not share common sites in BSA. Stern-Volmer analysis of fluorescence quenching data showed that the fraction of fluorophore (protein) accessible to the quencher (CB) was close to unity indicating thereby that both tryptophan residues of BSA are involved in drug-protein interaction. The rate constant for quenching, greater than $10^{10}$ $M^{-1}$ $s^{-1}$, indicated that the drug binding site is in close proximity to tryptophan residue of BSA. Thermodynamic parameters obtained from data at different temperatures showed that the binding of CB to BSA involves hydrophobic bonds predominantly. Significant increase in concentration of free drug was observed for CB in presence of paracetamol. Circular dichroism studies revealed the change in helicity of BSA due to binding of CB to BSA.

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Fluorescence probe study on the solubilization sites of aniline derivatives in triton X-100 and zephiramine micelles

  • Han, Suk-Kyu;Lee, Yong-Soo
    • Archives of Pharmacal Research
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    • v.9 no.3
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    • pp.139-144
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    • 1986
  • The solubilization sites of aniline and its derivatives in micelles were investigated with fluorescence probe technique. The fluorescence probes employed in this study are 12-(9-anthroyl) stearic acid (AS) and 2-p-toluidinylnaphthalene-6-sulfonate (TNS) which are incorporated in the interior of the micelle and attaced to its surface, respectively. As these two probes were effectively quenched by aniline and its surface, respectively. As these two probes were effectively quenched by aniline and its derivatives, the modified Stern-Volmer relationship in micellar system could be applicable to estimate the partition coefficient, $K_{p}$ of the solubilizate between aqueous and micellar phase. Because $K_{p}$ derived by this method reflects the relative proximity of the fluorophore to the quencher, the ratio of $K_{p}$ in the surface area to that in the interior of the micelle is interpreted in terms of the relative location of the solubilizate in micellar aggregate. The results show that the solubilizates are not located in a definite position but distributed in the multiple-sites of the micelle. The solubilization sites of the solubilizates in the icelle are dependent on their structures. As the solubilizate has more numbers of N-substituents of aniline and more numbers, of carbon in the substituent, it tends to incorporate in the interior of the micelle more effectively.

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Numerical Model for Thermal Hydraulic Analysis in Cable-in-Conduit-Conductors

  • Wang, Qiuliang;Kim, Kee-Man;Yoon, Cheon-Seog
    • Journal of Mechanical Science and Technology
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    • v.14 no.9
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    • pp.985-996
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    • 2000
  • The issue of quench is related to safety operation of large-scale superconducting magnet system fabricated by cable-in-conduit conductor. A numerical method is presented to simulate the thermal hydraulic quench characteristics in the superconducting Tokamak magnet system, One-dimensional fluid dynamic equations for supercritical helium and the equation of heat conduction for the conduit are used to describe the thermal hydraulic characteristics in the cable-in-conduit conductor. The high heat transfer approximation between supercritical helium and superconducting strands is taken into account due to strong heating induced flow of supercritical helium. The fully implicit time integration of upwind scheme for finite volume method is utilized to discretize the equations on the staggered mesh. The scheme of a new adaptive mesh is proposed for the moving boundary problem and the time term is discretized by the-implicit scheme. It remarkably reduces the CPU time by local linearization of coefficient and the compressible storage of the large sparse matrix of discretized equations. The discretized equations are solved by the IMSL. The numerical implement is discussed in detail. The validation of this method is demonstrated by comparison of the numerical results with those of the SARUMAN and the QUENCHER and experimental measurements.

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