• Title/Summary/Keyword: binding constant

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Brain Succinic Semialdehyde Dehydrogenase; Reaction of Arginine Residues Connected with Catalytic Activities

  • Bahn, Jae-Hoon;Lee, Byung-Ryong;Jeon, Seong-Gyu;Jang, Joong-Sik;Kim, Chung-Kwon;Jin, Li-Hua;Park, Jin-Seu;Cho, Yong-Joon;Cho, Sung-Woo;Kwon, Oh-Shin;Choi, Soo-Young
    • BMB Reports
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    • v.33 no.4
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    • pp.317-320
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    • 2000
  • The succinic semialdehyde dehydrogenase from bovine brain was inactivated by treatment with phenylglyoxal, a reagent that specifically modifies arginine residues. The inhibition at various phenylglyoxal concentrations shows pseudo-first-order kinetics with an apparent secondorder rate constant of 30 $M^{-1}min^{-1}$ for inactivation. Partial protection against inactivation was provided by the coenzyme $NAD^+$, but not by the substrate succinic semialdehyde. Spectrophotometric studies indicated that complete inactivation of the enzyme resulted from the binding of 2 mol phenylglyoxal per mol of enzyme. These results suggest that essential arginine residues, located at or near the coenzyme-binding site, are connected with the catalytic activity of brain succinic semialdehyde dehydrogenase.

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Postnatal Expression Pattern of Adipose Type Fatty Acid Binding Protein in Different Adipose Tissues of Porcine

  • Xu, C.L.;Wang, Y.H.;Huang, Y.H.;Liu, J.X.;Feng, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.811-816
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    • 2007
  • Adipocyte fatty acid-binding protein (A-FABP), which belongs to the FABP family, plays an essential role in long-chain fatty acid uptake and metabolic homeostasis, especially in adipose tissue. The pattern of A-FABP gene mRNA expression in different growth stages and its relation to intramuscular fat (IMF) accretion in pigs was studied. Fifteen female $Duroc{\times}Landrace{\times}Yorkshire$ pigs in five groups of three pigs each, weighing 1, 30, 50, 70 and 90 kg were used to study developmental gene mRNA expression of A-FABP in various adipose tissues by means of semi-quantitative RT-PCR. Results showed that A-FABP mRNA levels in subcutaneous and ventral adipose tissues first increased from 1 to 50 kg, then gradually declined from 50 to 90 kg. Moreover, the rank order of A-FABP mRNA levels determined in three adipose tissues was as follows: subcutaneous adipose>ventral adipose>mesenteric adipose. A-FABP mRNA expression in mesenteric adipose tissue was constant during development. In addition, a positive correlation from 1 to 50 kg BW pigs and a negative correlation from 50 to 90 kg BW between A-FABP mRNA levels in subcutaneous and ventral adipose and IMF content were found.

Photochemical Property and Photodynamic Activity of Tetrakis(2-naphthyl) Porphyrin Phosphorus(V) Complex

  • Hirakawa, Kazutaka;Aoki, Shunsuke;Ueda, Hiroyuki;Ouyang, Dongyan;Okazaki, Shigetoshi
    • Rapid Communication in Photoscience
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    • v.4 no.2
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    • pp.37-40
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    • 2015
  • To examine the photosensitized biomolecules damaging activity, dimethoxyP(V)tetrakis(2-naphthyl)porphyrin (NP) and dimethoxyP(V)tetraphenylporphyrin (PP) were synthesized. The naphthyl moiety of NP hardly deactivated the photoexcited P(V)porphyrin ring in ethanol. In aqueous solution, the naphthyl moiety showed the quenching effect on the photoexcited porphyrin ring, possibly through electron transfer and self-quenching by a molecular association. Binding interaction between human serum albumin (HSA), a water soluble protein, and these porphyrins could be confirmed by the absorption spectral change. The apparent association constant of NP was larger than that of PP. It is explained by that more hydrophobic NP can easily bind into the hydrophobic pockets of HSA. The photoexcited PP effectively induced damage of the tryptophan residue of HSA, through electron transfer-mediated oxidation and singlet oxygen generation. NP also induced HSA damage during photo-irradiation and the contributions of the electron transfer and singlet oxygen mechanisms were speculated. The electron transfer-mediated mechanism to the photosensitized protein damage should be advantageous for photodynamic therapy in hypoxic condition. The quantum yield of the HSA photodamage by PP was significantly larger than that of NP. The quenching effect of the naphthyl moiety is considered to suppress the photosensitized protein damage. In conclusion, the naphthalene substitution to the P(V)porphyrins can enhance the binding interaction with hydrophobic biomacromolecules such as protein, however, this substitution may reduce the photodynamic effect of P(V)porphyrin ring in aqueous media.

Thermodynamic Study on the Micellar Properties of DBS/Brij 30 Mixed Surfactant Systems (DBS/Brij 30 혼합계면활성제의 미셀화에 대한 열역학적 연구)

  • Lee, Byeong-Hwan;Park, In-Jeong
    • Journal of the Korean Chemical Society
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    • v.50 no.3
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    • pp.190-195
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    • 2006
  • The critical micelle concentrations (CMC) and the counter ion binding constants (B) in a micellar state of the mixed surfactant systems of sodium dodecylbenzenesulfonate (DBS) with polyoxyethylene(4) lauryl ether (Brij 30) in water were determined as a function of 1 (the overall mole fraction of DBS) by the use of electric conductivity method and surface tensiometer method from 288 K to 308 K. Various thermodynamic parameters (Smo, Hmo, and Gmo) for the micellization of DBS/Brij 30 mixtures were calculated and analyzed from the temperature dependence of CMC values. The measured values of Gomare all negative but the values of Smo are positive in the whole measured temperature region. On the other hand, the values of Hmo are positive or negative, depending on the measured temperature and 1.

Synthesis, Characterization and DNA Interaction Studies of (N,N'-Bis(5-phenylazosalicylaldehyde)-ethylenediamine) Cobalt(II) Complex

  • Sohrabi, Nasrin;Rasouli, Nahid;Kamkar, Mehdi
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2523-2528
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    • 2014
  • In the present study, at first, azo Schiff base ligand of (N,N'-bis(5-phenylazosalicylaldehyde)-ethylenediamine) ($H_2L$) has been synthesized by condensation reaction of 5-phenylazosalicylaldehyde and ethylenediamine in 2:1 molar ratio, respectively. Then, its cobalt complex (CoL) was synthesized by reaction of $Co(OAc)_2{\cdot}4H_2O$ with ligand ($H_2L$) in 1:1 molar ratio in ethanol solvent. This ligand and its cobalt complex containing azo functional groups were characterized using elemental analysis, $^1H$-NMR, UV-vis and IR spectroscopies. Subsequently, the interaction between native calf thymus deoxyribonucleic acid (ct-DNA) and CoL complex was investigated in 10 mM Tris/HCl buffer solution, pH = 7 using UV-vis absorption, thermal denaturation technique and viscosity measurements. From spectrophotometric titration experiments, the binding constant of CoL complex with ct-DNA was found to be $(2.4{\pm}0.2){\times}10^4M^{-1}$. The thermodynamic parameters were calculated by van't Hoff equation.The enthalpy and entropy changes were $5753.94{\pm}172.66kcal/mol$ and $43.93{\pm}1.18cal/mol{\cdot}K$ at $25^{\circ}C$, respectively. Thermal denaturation experiments represent the increasing of melting temperature of ct-DNA (about $0.93^{\circ}C$) due to binding of CoL complex. The results indicate that the process is entropy-driven and suggest that hydrophobic interactions are the main driving force for the complex formation.

Hypocholesterolemic Soybean Peptide (IAVP) Inhibits HMG-CoA Reductase in a Competitive Manner

  • Pak, Valeriy V.;Koo, Min-Seon;Lee, Na-Ri;Oh, Su-Kyung;Kim, Myung-Sunny;Lee, Jong-Soo;Kwon, Dae-Young
    • Food Science and Biotechnology
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    • v.14 no.6
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    • pp.727-731
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    • 2005
  • Synthesized Ile-Ala-Val-Pro (IAVP) peptide, which has the highest hypocholesterolemic effect among a number of synthesized derivatives of Ile-Ala-Val-Pro-Gly-Glu-Val-Ala (IAVPGEVA) isolated from 11S globulin of soy protein by pepsin digestion, was selected for investigation in the present study. Using a recombinant Syrian hamster 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), we studied in detail the inhibition of this enzyme by IAVP and compared the action of this peptide to that of lovastatin, a known competitive inhibitor of this enzyme. The concentration of IAVP required for 50% inhibition ($IC_{50}$) of HMGR activity in given experimental conditions was $340\;{\mu}M$. Kinetic analysis revealed that the studied peptide is a competitive inhibitor of HMGR with respect to both 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) and nicotinamide adenine dinucleotide phosphate (NADPH), with an equilibrium constant of inhibitor binding ($K_i\;=\;[E][I]/[EI]$) of $61{\pm}1.2\;{\mu}M$ and $157{\pm}4.4\;{\mu}M$, respectively. At the same conditions, $K_i$ and $IC_{50}$ for lovastatin were $2.2{\pm}0.1\;nM$ and 12.5 nM, respectively. Thus, the given peptide interacts with HMGR as a bisubstrate, consequently blocking access of both substrates to the active sites. The achieved results suggest the design of new peptide sequences having a higher relative affinity to binding sites of this enzyme and an enhancement of their hypocholesterolemic properties.

Percutaneous Absorption of Recombinant h-EGF through Normal, Stripped and First-Degree Burn Skin (상피세포 성장인자의 경피흡수: 정상피부, 각질제거피부 및 화상피부에 있어서)

  • Cho, Ae-Ri;Lee, Jung-Uk;Ahn, Byung-Lak;Chung, Joo-Young;Koh, Yeo-Wook;Shim, Chang-Koo
    • Journal of Pharmaceutical Investigation
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    • v.26 no.1
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    • pp.29-32
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    • 1996
  • In vivo and in vitro skin permeation of $recombinant^{125}$ I-EGF through normal, stripped and the first degree burn skin were studied. The in vitro skin permeation rate through the first degree burn skin $(296\;cpm/cm^2/hr)$ and the stripped skin $(1131\;cpm/cm^2/hr)$ were 3.5 times and 13 times higher, respectively, as compared with the one through normal skin. In vivo absorption study with the first degree burn skin, the peak concentration of EGF in the skin was achieved at 1-3 hr and decreased afterward up to 8 hr with an elimination constant of $1.31{\times}10^{-3}\;g/ml/hr$. To investigate the higher elimination rate of EGF in burn skin, binding and metabolism studies were conducted. No significant metabolism of EGF in burn skin $(100^{\circ}C,\;5-second\;burning)$ was observed. With the presence or unlabelled-EGF $^{125}I-EGF$ permeation through the burn skin showed higher permeation rate than the one without unlabelled-EGF. The result nay indicate that EGF-receptor binding play a role in determining the skin permeation rate.

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Formation of a Unique 1:2 Calcium-Calixquinone Complex in Aqueous Media

  • Kang, Sun-Kil;Lee, One-Sun;Chang, Suk-Kyu;Chung, Doo-Soo;Kim, Ha-Suck;Chung, Taek-Dong
    • Bulletin of the Korean Chemical Society
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    • v.32 no.3
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    • pp.793-799
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    • 2011
  • We report the complexation behavior of calix[4]arenemonoquinone-triacid (CTAQ), which is an electroactive and water-soluble receptor for calcium ion. UV-visible and NMR spectroscopic studies revealed that CTAQ in aqueous media forms 1:2 as well as 1:1 (metal ion:CTAQ) stoichiometric complexes with $Ca^{2+}$, $Sr^{2+}$, and $Ba^{2+}$ ions. The nonlinear fitting of titration curves based on UV-visible absorption spectra showed that the binding constants of CTAQ for $Ca^{2+}$ ion are 4 $({\pm}2){\times}10^6\;M^{-1}$ for 1:1 and 1.4 $({\pm}0.5){\times}10^{11}\;M^{-2}$ for 1:2 complex. NMR conformational studies and the titration curves corroborate that the $Ca^{2+}$:CTAQ complex in aqueous solution is not present in the form of merely 1:1 one, being consistent with UV-visible spectrophotometric results. The Monte Carlo simulation supports the presence of a stable conformer of 1:2 complexes in which a $Ca^{2+}$ ion is interposed between two CTAQs at the global minimum. This is the first model of 1:2 stoichiometric complex of calix[4]arene and alkaline earth ions in aqueous media.

Development of a Fluoride-Selective Electrode based on Scandium(III) Octaethylporphyrin in a Plasticized Polymeric Membrane

  • Kang, Young-Jea;Lutz, Christopher;Hong, Sung-A;Sung, Da-Yeon;Lee, Jae-Seon;Shin, Jae-Ho;Nam, Hak-Hyun;Cha, Geun-Sig;Meyerhoff, Mark E.
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1601-1608
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    • 2010
  • A scandium(III) porphyrin-based fluoride-selective potentiometric sensor and its application in the analysis of hydrofluoric acid is described. Scandium(III) octaethylporphyrin, an ionophore recently developed for the optical fluoride sensor, was employed as a host molecule for the selective binding with fluoride in the plasticized PVC membrane. Nernstian response for $F^-$ between $10^{-4.6}$ to $10^{-1}$ M was observed at a glycine-phosphate buffer (pH 3.0). The selectivity pattern was observed as $F^-$, salicylate $\gg$ $SCN^-$ > $Cl^-$, $Br^-$, $NO_3{^-}$, $ClO_4{^-}$, which is consistent with the binding constant data measured in the plasticized PVC membrane based on a sandwich membrane method. This highly selective and reversible fluoride-sensitive electrode was employed for the analysis of hydrofluoric acid (HF). A disposable differential-type HF sensor was fabricated on the screen-printed electrode and demonstrated its ability to detect the neutral HF in the acidic solution.

Determination of inclusion complex formation constants for the β-CD and [Cu(Dien)(sub-Py)]2+ ion by the spectrophotometric methods (분광 광도법에 의한 β-CD와 [Cu(Dien)(sub-Py)]2+이온간의 복합체 형성 상수 결정)

  • Kim, Chang Suk;Oh, Ju Young
    • Analytical Science and Technology
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    • v.20 no.5
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    • pp.406-412
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    • 2007
  • The formation of inclusion complexes between ${\beta}$-cyclodextrin and diethylenetriamine substituted-pyridine copper(II) perchlorate; [Cu(dien)(sub-py)] $(ClO_4)_2$, were studied by spectrophotometric methods. On account of charge-transfer band(MLCT) and $^2T_2{\rightarrow}^2E$, the two high peaks were observed as an inclusion complex for the [${\beta}$-CD]$[Cu(dien)(p-Cl-py)]^{2+}$ in the ultraviolet region of the spectrum. The ${\beta}$-CD and $[Cu(dien)(sub-py)]^{2+}$ ion formed a 1:1 complex, and the formation constants were decreased with the increasing temperatures, due to weak binding energy between ${\beta}$-CD and $[Cu(dien)(sub-py)]^{2+}$ ion. This reaction was controlled by enthalpy. In a correlation of the Hammett substituent constants and formation constants for the reaction, formation constants were increased by strong binding energy in the inclusion complexes when electron donating groups were substituted in pyridine ring.