• Title/Summary/Keyword: binding constant(K)

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Interaction of ${\alpha}$-Ketoglutarate Dehydrogenase Complex with Allosteric Regulators Detected by a Fluorescence Probe, 1,1'-bi(4-aniline)naphthalene-5,5'-disulfonic acid, an Inhibitor of Catalytic Activity

  • Hong, Sung-Youl;Bak, Choong-Il;Ryu, Jae-Ha;Song, Byoung-J.;Huh, Jae-Wook
    • BMB Reports
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    • v.29 no.3
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    • pp.230-235
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    • 1996
  • The interaction of ${\alpha}-ketoglutarate$ dehydrogenase complex (${\alpha}-KGDC$) with a hydrophobic fluorescent probe [1,1'-bi(4-aniline)naphthalene-5,5'-disulfonic acid] (bis-ANS) was studied. The punfied ${\alpha}-KGDC$ was potently inhibited by bis-ANS with an apparent half maximal inhibitory concentration ($IC_{50}$) of 9.8 ${\mu}m$ at pH 8.0. The catalytic activities of both the E1o and E2o subunits were predominantly inhibited while that of the E3 component was hardly affected. The binding of bis-ANS to the enzyme caused a marked enhancement and blue shift from 523 nm to 482 nm in the fluorescence emission spectrum. The dissociation constant ($K_d$) and the number of binding sites (n) were calculated to be 0.87 mM and 158, respectively. Allosteric regulators such as purine nucleotides and divalent cations further increased the fluorescence intensity of the $bis-ANS-{\alpha}-KGDC$ binary complex. These data suggest that the binding of these allosteric regulators to ${\alpha}-KGDC$ may cause the conformational changes in the enzyme and that bis-ANS could be used as a valuable probe to study the interaction of the multi-enzyme complex and its allosteric regulators.

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Structural, Electrochemical, DNA Binding and Cleavage Properties of Nickel(II) Complex [Ni(H2biim)2(H2O)2]2+ of 2,2'-Biimidazole

  • Jayamani, Arumugam;Thamilarasan, Vijayan;Ganesan, Venketasan;Sengottuvelan, Nallathambi
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3695-3702
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    • 2013
  • A nickel(II) complex $[Ni(H_2biim)_2(H_2O)_2](ClO_4)_2{\cdot}H_2O$ (1) of biimidazole ligand has been synthesized and characterized (Where $H_2biim$ = 2,2'-biimidazole). The single crystal X-ray diffraction of the complex shows a dimeric structure with six coordinated psudo-octahedral geometry. The cyclic voltammograms of complex exhibited one quasireversible reduction wave ($E_{pc}=-0.61V$) and an irreversible oxidation wave ($E_{pa}=1.28V$) in DMF solution. The interaction of the complex with Calf-Thymus DNA (CT-DNA) has been investigated by absorption and fluorescence spectroscopy. The complex is an avid DNA binder with a binding constant value of $1.03{\times}10^5M^{-1}$. The results suggest that the nickel(II) complex bind to CT-DNA via intercalative mode and can quench the fluorescence intensity of EB bind to CT-DNA with $K_{app}$ value of $3.2{\times}10^5M^{-1}$. The complex also shown efficient oxidative cleavage of supercoiled pBR322 DNA in the presence of hydrogen peroxide as oxidizing agent. The DNA cleavage by complex in presence of quenchers; viz. DMSO, KI, $NaN_3$ and EDTA reveals that hydroxyl radical or singlet oxygen mechanism is involved. The complex showed invitro antimicrobial activity against four bacteria and two fungi. The antimicrobial activity was nearer to that of standard drugs and greater than that of the free ligand.

Separation of Cu(II) from Metal Mixture Solution Using a Hexadecyl $NtnOenH_4$-Octadecylsilanized Silicas(ODS) in Liquid Chromatography (액체 크로마토그래피에서 Hexadecyl $NtnOenH_4$-Octadecylsilanized silicas(ODS)를 이용한 혼합금속용액으로부터 Cu(II)의 분리)

  • Shin, Young-Kook;Kim, Si-Joong;Kim, Hae-Joong
    • Analytical Science and Technology
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    • v.8 no.3
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    • pp.299-304
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    • 1995
  • The adsorption behaviors of Ba(II), Cr(II), Fe(II) and Cu(II) on the N, N'-bispalmtoyl 1, 12-diaza-3, 4:9, 10-dibenzo-5, 8-cyclopentadecane (hexadecyl $NtnOenH_4$)-octadecylsilanized silicas(ODS) were investigated with water as the mobile phase. Binding constants for metal ions were measured in aqueous solution. The order of binding constants(K) and the degree of sorption(E) were Ba(II)$NtnOenH_4$-octadecyisiianized silicas(ODS) increased with concentration of metal ions, and the degree of adsorption was found to be affected by the cation-chelation mechanism. The experimental results showed good efficiency for separation of Cu(II) from mixtures of Ba(II), Cr(II), Fe(II) and Cu(II) in aqueous solution.

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Binding Profiles of Oxomemazine to the Muscarinic Receptor Subtypes (Oxomemazine의 Muscarinic Receptor Subtypes에 대한 결합성질)

  • Lee, Shin-Woong;Kim, Jeung-Gu
    • The Korean Journal of Pharmacology
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    • v.30 no.1
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    • pp.49-57
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    • 1994
  • The binding properties of oxomemazine to muscarinic receptors using the ability of oxomemazine to inhibit $[^3H]QNB$ binding in membrane fractions of rat cerebrum and guinea pig ventricle and ileum were investigated. $[^3H]QNB$ bound to a single class of muscarinic receptors with a dissociation constant of approximately 60 pM in three tissue preparations. Pirenzepine and oxomemazine inhibited $[^3H]QNB$ binding in cerebrum with a Hill coefficient lower than unity, and the inhibition data were best described by a two-site model. The relative densities of the high $(M_1)\;and\;low\;(M_2)$ affinity sites for pirenzepine were 60 and 40%, with corresponding Ki values of 16 and 431 nM, and those $(O_H\;and\;O_L)$ for oxomemazine 40 and 60%, with corresponding Ki values of 80 and 1350 nM. However, the inhibition data of both drugs vs $[^3H]QNB$ in ventricle and ileum appeared to obey the law of mass-action (Hill coefficient close to 1). The apparent Ki values of pirenzepine were 850 and 250 nM, and those of oxomemazine 1460 and 570 nM in ventricle and ileum, respectively. Thus, oxomemazine like pirenzepine has high affinity for cerebrum, moderate affinity for ileum and low affinity for ventricle. These results suggest that oxomemazine could recognize the muscarinic receptor subtypes with a high affinity for the $M_1$ sites.

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Comparison of the Stability Constants of Cd(II)-, Cu(II)-, and Pb(II)-Humate Complexes

  • Choi, Se-Young;Moon, Hi-Chung;Jun, Song-Hui;Chung, Kun-Ho
    • Bulletin of the Korean Chemical Society
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    • v.15 no.7
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    • pp.581-584
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    • 1994
  • A comparative investigation of the complexations of divalent metal ions (Cd, Cu, and Pb) by a well characterized soil humic acid (HA) from Okchun Metamorphic Belt was carried out in 0.05 M $KNO_3$ and pH of 4.5 using ion selective electrodes. A continuous distribution model based on the Scatchard Plot was used to determine the stability constants, because the constants obtained by this modeling technique takes the variations in binding energies into consideration without regards to the manner in which M(II) ion is bound on HA. The mean value of log $K_i$ were $4.05{\pm}0.60,\;4.92{\pm}0.36,\;and\;5.63{\pm}0.34\;{\ell}\;mol^{-1}$ for Cd(II)-, Pb(II), and Cu(II)-humate complexes respectively. The values of intrinsic constant (log $K_{int}$; binding at strongest site) were $7.12{\pm}0.30,\;6.59{\pm}0.32,\;and5.07{\pm}0.56\;{\ell}\; mol^{-1}$ in the order Cu(II) > Pb(II) > Cd(II) ion.

Determination of Stability Constants of the Inclusion Complexes of ${\beta}$-Blockers in Heptakis (2,3-Dimethyl-6-Sulfato)- ${\beta}$-Cyclodextrin

  • Phuong, Nuyen Thi;Lee, Kyung-Ah;Kim, Kyung-Ho;Choi, Jung-Kap;Kim, Jong-Moon;Kang , Jong-Seong
    • Archives of Pharmacal Research
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    • v.27 no.12
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    • pp.1290-1294
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    • 2004
  • The ${\beta}$-blockers possess at least one chiral center and the S(-)-enantiomer shows higher affinity for binding to the ${\beta}$-adrenergic receptors than antipode. The stability constants of acebutolol, celiprolol, propranolol and terbutaline in the inclusion complexes with single-isomer heptakis (2,3-dimethyl-6-sulfato)- ${\beta}$-cyclodextrin (HDMS-${\beta}$-CD) were determined by capillary electrophoresis. The approximation and linear double reciprocal methods were adapted with comparable results. Among the ${\beta}$-blockers studied, propranolol had the lowest stability constant but the highest enantioselectivity, indicating that the magnitudes of the stability constants carried little information about enantioseparation. The magnitudes of enantioselectivities between the enantiomer pair were in the order of propranolol > celiprolol > terbutaline > acebutolol.

Effect of n-Butanol on the Mixed Micellization of DPC with CDEAB (DPC와 CDEAB의 혼합마이셀화에 미치는 n-부탄올의 효과)

  • Lee, Byung-Hwan
    • Journal of the Korean Chemical Society
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    • v.48 no.3
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    • pp.236-242
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    • 2004
  • The critical micelle concentration (CMC) and the counterion binding constant (B) in a mixed micellar state of the Dodecylpyridinium chloride (DPC) with the Cetyldimethylethylammonium bromide (CDEAB) at 25$^{\circ}C$ in aqueous solutions of n-butanol were determined as a function of ${\alpha}_1$ (the overall mole fraction of DPC) by the use of electric conductivity method. Various thermodynamic parameters (($X_i,\;{\gamma}_i,\;C_i,\;a^M_i,\;{\beta},\;and {\Delta}H_{mix})$were calculated by means of the equations derived from the nonideal mixed micellar model. The effect of n-butanol on the mixed micellization of the DPC/CDEAB mixtures has been also studied by analyzing the measured and calculated thermodynamic parameters (CMC, B 및 $;{\Delta}G_o\;^m$).

Enantioselective electrophoretic behavior of lipoic acid in single and dual cyclodextrin systems

  • Le, Thi-Anh-Tuyet;Nguyen, Bao-Tan;Phan, Thanh Dung;Kang, Jong-Seong;Kim, Kyeong Ho
    • Analytical Science and Technology
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    • v.34 no.4
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    • pp.143-152
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    • 2021
  • Capillary electrophoresis (CE) is an effective technique to study chiral recognition because it offers flexibility in adjusting vital factors. Currently, various available cyclodextrins (CDs) can be employed for the chiral separation of numerous analytes. Herein, we investigate the enantioseparation behavior of lipoic acid enantiomers in various types of single and dual CD systems through CE. Additionally, several impacted CE parameters were optimized through the systematic investigation based on the design of experiment (DoE) concept for a single system comprising a heptakis (2,3,6-tri-O-methyl)-β-CD and a dual system containing the combination of the single CD with a sulfated-β-CD. Consequently, absolute enantioresolution was obtained within 15 min on a common standard bare fused-silica capillary (64.5/56 cm in total/effective length, 50/365 ㎛ inner/outer diameter), maintained at 15 ℃ and at an applied voltage of 24 kV. The optimal background electrolyte consisted of 6 mM heptakis (2,3,6-tri-O-methyl)-β-CD dissolved in the solution of 58 mM borate buffer at pH 10. Furthermore, the results of apparent binding constant experiments indicated that the S-enantiomer-heptakis (2,3,6-tri-O-methyl)-β-CD complex exhibited a stronger affinity than its R-enantiomer counterpart. The obtained electrophoretic mobility values could be utilized to interpret the resolution achieved at various CD concentrations and the mobility behavior of the complexes elucidated the migration order of the enantiomers in an electropherogram.

Effect of Carbohydrates to Protein Ratio in EPS on Sludge Settling Characteristics

  • Shin, Hang-Sik;Kang, Seok-Tae;Nam, Se-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.460-464
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    • 2000
  • Extracellular polymeric substances (EPS) are believed to play a role in the binding and formation of microbial flocs. However, the precise role is not well known. Sludge settling characteristics and the carbohydrate to protein ratio in EPS were tested with various airflow rates in this study. Sludge was collected from three modified sequencing batch reactors (SBRs), which were operated at 16$\^{C}$ with an airflow rate of 0.8L/min, 3L/min and 6L/min, respectively. During the operation, the reactor operated at an airflow rate of 0.8L/min showed sludge volume index (SVI) of 80 to 90ml/g and a constant ratio of carbohydrate to protein in the EPS, while a significant increase in the SVI was seen in the other reactors. Sludge bulking increased the amount of carbohydrate in the EPS, while kept protein almost constant in the airflow rate of 3L/min ad 6L/min. Surface charge also increased with increases in the carbohydrate to protein ratio in the EPS, which weakens the attraction between the EPS and multivalent cations. The ratio of carbohydrate to protein in the EPS was tween the EPS and multivalent cations. The ratio of carbohydrate to protein in the EPS was inferred to be essential for bioflocculation.

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Sheet Reisistance Analysis of TiNx Thin Film by RF Magnetron Sputtering (RF magnetron 스파터링법으로 제작한 TiNx 박막의 면저항분석)

  • Park, Moon Chan;Oh, Jeong Hong;Kim, Nam Young;Hwangbo, Chang Kwon
    • Journal of Korean Ophthalmic Optics Society
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    • v.4 no.1
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    • pp.21-25
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    • 1999
  • The TiN, thin films were prepared on glass substrate by RF(radio-frequency) magnetron sputtering apparatus from a Ti target in a gaseous mixture of argon and nitrogen. In deposition, a RF power supply was used as a power source with a constant power of 240W, and the substrate was heated to $200^{\circ}C$. The films were obtained at nitogen flow rates in the range 3-9 sccm with a constant argon flow rate of 20 secm. For the films obtained, the sheet resistance and the chemical binding energy of the films was observed by four-point-probe method and x-ray photoelectron spectroscopy(XPS) depth profiling respectively. In addition, we investigated the relationship between the surface resistance and the chemical nature of the films.

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