• Title/Summary/Keyword: Carbodiimide

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Production and Characterization of Specific Antibodies to Bombesin

  • Kwon, Hyeok-Yil;Lee, Yun-Lyul;Park, Hyoung-Jin
    • The Korean Journal of Physiology
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    • v.28 no.1
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    • pp.91-97
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    • 1994
  • In order to produce a specific bombesin antiserum far very sensitive radioimmunoassay, synthetic $[lys^3]-bombesin$ conjugated to bovine serum albumin was subcutaneously injected into guinea pigs. The conjugation was performed using either carbodiimide or gIutaraldehyde as a coupling agent. The antisera were characterized by analysis of Scatchard and Sips plots. The antiserum LBE 2G/2 raised by repeat injection of the immunogen conjugated with carbodiimide showed the titer of 1 : 188,000, very low cross-reactivity to bombesin-like peptides except bombesin, with high affinity constant $(1.64{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.91). The antiserum LBG 1G/2 produced by repeat injection of the immunogen conjugated with glutaraldehyde possessed the titer of 1 : 43,000, high cross-reactivity to some bombesin-like peptides, high affinity constant $(1.19{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.79). These results indicate that the antiserum LBE 2G/2 is specific only to bombesin and that the antiserum LBG IG/2 binds to some bombesin-like peptides such as alytesin, gastrin releasing peptide and neuromedin C. The antiserum LBE 2G/2 is sufficient for the very sensitive radioimmunoassay of bombesin.

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Purification and Characterization of 5,10-Methenyltetrahydrofolate Synthetase from Chicken Liver (닭의 간 유래의 5,10-Methenyltetrahydrofolate Synthetase의 정제 및 특성)

  • Cho, Yong-Kweon
    • Journal of the Korean Chemical Society
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    • v.54 no.5
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    • pp.567-572
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    • 2010
  • 5,10-Methenyltetrahydrofolate synthetase from chicken liver was purified through 30-70% ammonium sulfate fractionation, Q Sepharose Fast Flow anion exchange and Source 15Phe hydrophobic interaction chromatography. Specific activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 0.0085, 0.031, 0.80 and 1.27 U/mg, respectively. Purification fold activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 1, 3.7, 94.1 and 149.4, respectively. HPLC gel permeation chromatography and SDS-polyacrylamide electrophoresis experiments indicated that the enzyme is a monomeric protein with a molecular weight of 22.8 kDa. Km for 5-methyl THF and Mg-ATP were $7.1\;{\mu}M$ and $63\;{\mu}M$, respectively. Optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. The data for metal ion specificity and stoichiometry showed that the maximum activity was obtained with a 1:l. ratio of $Mg^{2+}$. The ATP and Km values increased in the order of MgATP, MgCTP, MgUTP and MgGTP, and the maximum activities also decreased in the same order, indicating MgATP as the most efficient substrate. The enzyme was chemically modified only by tetranitrometane and 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, indicating that tyrosine and carboxylate are present in the active site.

Immunological Studies on the Natural Products I -Production of Antibody Specific to Saikosaponin a-

  • Sung, Chung-Ki
    • Korean Journal of Pharmacognosy
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    • v.19 no.2
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    • pp.127-132
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    • 1988
  • In the course of the immunological studies on the natural products, the antibody specific to saikosaponin a was producted. Saikosaponin a was treated with succinic anhydride to give 6'-O-hemisuccinyl saikosaponin a, which was successively converted to saikosaponin a-BSA conjugate (4. 5 mole saikosaponin a/mole of BSA) by carbodiimide method. The antibody obtained from rabbits immunized with saikosaponin a-BSA conjugate as usual manner reacted with both the conjugate and BSA, while after the absorption with BSA, the antibody reacted with the conjugate alone.

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Heterocyclization Reaction of 4-(2-Methylaziridin-1-yl)-3-ureidobenzotrifluorides under Appel's Conditions

  • Cho, Hyun-In;Lee, Kee-Jung
    • Bulletin of the Korean Chemical Society
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    • v.24 no.2
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    • pp.189-192
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    • 2003
  • The reaction of 4-(2-methylaziridin-1-yl)-3-ureidobenzotrifluorides 4 with triphenylphosphine, carbon tetrachloride, and triethylamine (Appel's condition) led to the corresponding carbodiimides 5, which underwent intramolecular cycloaddition reaction with aziridine under the reaction condition to give the benzimidazolefused heterocycles, 2,3-dihydro-1H-imidazo[1,2-a]benzimidazoles 8 and 12,13-dihydro-5H-benzimidazo[2,3-b] [1,3]benzodiazepines 9.

Synthesis and Antiinflammatory-analgesic Activity of Cinmetacin Amides (Cinmetacin의 amide 유도체 합성과 항염진통활성)

  • 임채욱;홍용기;임철부
    • YAKHAK HOEJI
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    • v.45 no.6
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    • pp.565-569
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    • 2001
  • Five novel cinmetacin amide derivatives as potential nonsteroidal antiinflammatory and analgesic compounds were prepared and their antiinflammatory-analgesic activity was compared with cinmetacin. Cinmetacin and hydroxysuccinimide were reacted with dicyclohexyl carbodiimide to give cinmetacin active ester (4), which was treated with amines to yield cinmetacin amides (5-9). Compounds (5) and (9) gave stronger analgesic activity than cinmetacin and compounds (5), (6), (9) showed comparable antiinflammatory activity to cinmetacin.

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Synthesis and Analgesic-antiinflammatory Activity of Cinmetacin Derivatives (Cinmetacin유도체의 합성과 진통항염활성)

  • 임채욱;이종민;유재학;이현수;임철부
    • YAKHAK HOEJI
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    • v.45 no.1
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    • pp.1-6
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    • 2001
  • Nine cinmetacin derivatives as potential nonsteroidal analgesic and antiinflammatory compounds were prepared and their analgesic-antiinflammatory activity was compared with cinmetacin. Salicylic acid and phenols were reacted with dicyclohexyl carbodiimide (DCC) to give phenol salicylates (1-4). Cinmetacin was treated with DCC and phenol derivatives to yield cinmetacin esters (5-13). Compounds (5, 7, 8, 10, 12, and 13) showed stronger analgesic activity than cinmetacin, but only compound (5) showed comparable antiinflammatory activity to cinmetacin.

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Synthesis of 2,7-Methano-aza[10]annulene Derivatives

  • 김충섭
    • Bulletin of the Korean Chemical Society
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    • v.17 no.5
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    • pp.437-442
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    • 1996
  • Electrocyclic ring-closure of 6-vinylcyclohepta-1,3,5-isocyanate has been carried out in the presence of triphenylphosphine to examine a catalyzing effect of the triphenylphosphine. The preparation of 10-(1-carboalkoxyalkyl)-2,7-methanoaza[10]annulenes by the electrocyclic ring-closure of ketenimine intermediates, which are formed by the reaction of triphenylalkylidenephosphorane and 6-vinylcyclo-hepta-1,3,5-isocyanate, is described. 10-Alkyl-2,7-methanoaza[10]annulenes were prepared by basic hydrolysis of the carboalkoxyaza[10]annulenes and decarboxylation of the acid intermediates. In the same manner, 10-(N-alkyl(or aryl))-2,7-methanoaza[10]annulenes were prepared from the reaction of the isocyanate and N-alkyl(or aryl)iminotriphenylphosphorane via electrocylic ring-closure of carbodiimide intermediate.

Characterization of Hyaluronic Acid Membrane Cross-linked with Lactide (락타이드로 가교시킨 히아루론산 막의 특성)

  • Kwon, Ji-Young;Cheong, Seong-Ihl
    • Polymer(Korea)
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    • v.29 no.6
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    • pp.599-604
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    • 2005
  • The hyaluronic acid (HA) with excellent biocompatibility has been combined with lactide, the ester dimer of polylactide, with good biodegradability to produce biocompatible materials which can control the period of degradation in a human body. By freeze frying method, HA and lactide were crosslinked with crosslinking agent, 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC). Degree of lactide and EDC reaction was determined by the analysis of nuclear magnetic resonance spectroscopy. Both lactyl group and EDC conversion increased as the mole ratio of lactide to HA increased from 5 to 13. The membrane swelled less and became more brittle with the more addition of lactyl group resulting from the higher mole ratio of lactide to HA. Swelling ratio decreased and tensile modulus increased due to the more addition of lactyl group as the EDC concentration increased or reaction temperature decreased. Drug release experiment from various membranes with different degree of crosslinking showed that permeability decreased with increasing degree of crosslinking. The degradation became slower with the more addition of lactyl group. Mechanical property and degradation rate of the synthesized membrane were shown to be controlled through adjusting operation parameters such as mole ratio, temperature, and crosslinking agent concentration.

Kinetic and Chemical Mechanism of Pig Liver Methenyltetrahydrofolate Synthetase (돼지 간 유래의 methenyltetrahydrofolate synthetase 반응메커니즘)

  • Cho, Yong-Kweon
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1036-1041
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    • 2008
  • Methenyltetrahydrofolate synthetase extract was obtained from mouse liver and purified via $30{\sim}70%$ ammonium sulfate fractionation, Fast Q anion exchange and phenyl agarose chromatography. HPLC gel chromatography and SDS-polyacrylamide electrophoresis experiments showed that the enzyme is a monomer with molecular weight of 23 kDa. Optimum temperature and pH were $35^{\circ}C$ and 6.5, respectively. The enzyme was chemically modified only by tetranitromethane and 1-ethyl-3- (3-dimethyl aminopropyl)-carbodiimide (EDC), indicating that tyrosine and carboxylate are in the active site. pH studies showed that 2 tyrosines are involved in the binding of the substrates and a carboxylate in catalysis. Therefore, the chemical mechanism of the enzyme is likely that 2 tyrosines bind to ATP and 5-formylTHFand a carboxylate acts as a general base.

Poly-3,4-dihydroxybenzoic Acid Film Electrodes Modified with Dopamine for Determination of Ti(IV) Ions (도파민으로 수식된 3,4-dihydroxybenzoic acid 고분자 피막전극을 이용한 Ti(IV)이온의 정량)

  • Cha, Seong-Keuck
    • Journal of the Korean Electrochemical Society
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    • v.6 no.2
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    • pp.130-133
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    • 2003
  • 3,4-dihydroxybenzoic acid(3,4-DHBA) was electropolymerized on glassy carbon electrode to give the GC/p-3,4-DHBA type electrode which was modified with dopamine by the help of 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride(EDC) acting as a coupling agent. The carboxylic sites on the polymeric surface of p-3,4-DHBA and mine group at the dopamine gave a QCA(Au)/p-3,4-DHBA-dopamine type of modified electrodes. The o-quinone moieties at the electrode surface exhibited high selectivity to titanium ions in solution. The redox process of the electrode is hydroquinone : quinone +$2H^+2e^-$, which had two strong and two weak pairs of peaks at CV. The modified electrode can deposit Ti(IV) ions as much as $4.13\times10^{-5}gcm^{-2}$. The calibration curve of the electrodes, log of the surface coverage-normalized redox response vs log[Ti], exhibited an excellent correlation$(r{\geq}0.997)$ for titanium concentrations ranging from $5.25\times10^{-4}\;to\;5.25\tiems10^{-8}M.$.