• Title/Summary/Keyword: positively charged

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Characterization of Cysteine Residues in Cabbage Phospholipase D by Sulfhydryl Group Modifying Chemicals (설프히드릴 변형 화합물질들에 의한 양배추 포스포리파제 D의 시스테인 잔기의 특성)

  • Go, Eun-Hui
    • Journal of the Korean Chemical Society
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    • v.50 no.5
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    • pp.362-368
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    • 2006
  • SH group modifying chemicals were used to characterize the eight cysteine residues of cabbage PLD. 5,5-dithiobis(2-nitrobenzoate)(DTNB) was used to titrate the SH group of cysteine residues . Based on the optical density at 412nm due to the reduced DTNB, 4 SH groups are found to be present in a native PLD while 8 SH groups in the denatured PLD whose tertiary structure was perturbed by 8M urea. The results imply that among the 8 cysteine residues of PLD, the half(4) are exposed on the surface whereas the other half are present at the interior of the enzyme tertiary structure. The PLD was inactivated by SH modifying reagents such as p-chloromercuribenzoate(PCMB), iodoacetate, iodoacetamide, and N-ethylmaleimide. At the addition of dithiothreitol(DTT) only the PCMB inhibited PLD activity was recovered reversibly. The micro-environment of the exposed SH group of cysteine residues was examined with various disulfide compounds with different functional groups and we found that anionic or neutral disulfides appear to be more effective than the positively charged cystamine for inactivating the PLD activity. The effect of redox state of cysteine residues on the PLD activity was further explored with H2O2. The oxidation of SH groups by H2O2 inhibited the PLD activity more than 70%, which was mostly recovered by DTT. From these results, we could confirm chemically that all the cysteine residues of PLD are present as in their reduced SH forms and the 4 SH groups exposed on the surface of the enzyme may play important roles in the regulation of PLD activity.

The Effect of PDGF-Loaded Biodegradable Membrane on Early Healing Stage in Guided Tissue Regeneration (흡수성 차폐막의 치주조직 재생에 혈소판유래 성장인자가 미치는 영향)

  • Rhyu, In-Chul;Bae, Kyoo-Hyun;Seol, Yang-Jo;Ku, Young;Lee, Seung-Jin;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.29 no.3
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    • pp.507-519
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    • 1999
  • The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect f a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/$cm^2$ platelet derived growth factor. 2 beagle dogs were used in he experiment. $5mm{\times}6mm$ alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test roup or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new one formation could be seen. Small amount f bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane nd the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.

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Response Mechanism of 5, 10, 15, 20-tetraphenyl(porphyrinato) Manganase(III) chloride-Based Ion-Selective Membranes (망간포르피린을 함유한 고분자형 이온선택성 막전극의 감응 메카니즘)

  • Hong, Young Ki;Kang, You Ra;Shin, Dae Ho;Shin, Doo Soon;Cha, Geun Sig;Nam, Hakhyun
    • Analytical Science and Technology
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    • v.9 no.3
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    • pp.270-278
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    • 1996
  • Response mechanism for the chloride-selective membrane doped with 5, 10, 15, 20-tetraphenyl(porphyrinato)manganase(III) chloride(Mn(TPP)Cl) in PVC/DOS matrix is proposed by examining the visible spectra of the corresponding optode membrane. The visible spectra of Mn(TPP)Cl-doped membrane placed in aqueous solution show that the chloride ligand is easily replaced with water molecule. When other halogen ions, such as $F^-$, $Br^-$ and $I^-$, are added to the sample solution, they replace the water ligand, exhibiting distinctive change in the Soret band of Mn(TPP). On the other hand, bulky anions, such as SCN and salicylate, do not form a bond with the central metal. These results suggests that the potentiometric response of Mn(TPP)-based membrane results either from the ligand exchange (water with halides) at the central metal or from the counter ion exchange (chlorides with bulky lipophilic anions) around the positively charged porphyrin molecule in membrane phase. It was also noted that both hydration enthalpies of anions and their binding constants to Mn(TPP) play critical role in determining the potentiometric selectivity pattern of the membrane.

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Flotation Characteristics of Malachite by Various Collectors (포수제(捕收劑) 종류(種類)에 따른 공작석(孔雀石)의 부유선별(浮遊選別) 특성(特性))

  • Kim, Hyung-Seok;Kim, Wan-Tae;Han, In-Kyu;Kim, Sang-Bae
    • Resources Recycling
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    • v.19 no.4
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    • pp.58-64
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    • 2010
  • In order to select the proper collector for the flotation of malachite, we had floated malachite by using various collectors. When measuring the zeta potential of malachite, we knew that the point of zero charge of malachite is found at pH 8.2 and malachite particles are negatively charged above this pH value; and positively, below this pH value. The floatability of malachite by the anionic collectors like sodium oleate and Aeropromoter 845, was about 97% in pH 5~11 range, whereas that of malachite by alkyl hydroxamate and the cationic collector like dodecyl trimethyl ammonium chloride and dodecyl ethyl methyl ammonium chloride, was below 15% above pH 5 regardless of dosage increase of collector and flotation time. The malachite sulphidised by $(NH_4)_2S$ was floated above 97% by potassium amyl xanthate. The dosage of potassium amyl xanthate decreased and the flotation period shortened to one third than when it was not sulphidised by $(NH_4)_2S$. Therefore, potassium amyl xanthate and the anionic collectors like sodium oleate and Aeropromoter 845 can be used as the collector of malachite.

Contribution of Arginine 13 to the Catalytic Activity of Human Class Pi Glutathione Transferase P1-1

  • Kong, Ji-Na;Jo, Dong-Hyeon;Do, Hyun-Dong;Lee, Jin-Ju;Kong, Kwang-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.31 no.9
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    • pp.2497-2502
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    • 2010
  • Arg13 is a conserved active-site residue in all known Pi class glutathione S-transferases (GSTs) and in most Alpha class GSTs. To evaluate its contribution to substrate binding and catalysis of this residue, three mutants (R13A, R13K, and R13L) were expressed in Escherichia coli and purified by GSH affinity chromatography. The substitutions of Arg13 significantly affected GSH-conjugation activity, while scarcely affecting glutathione peroxidase or steroid isomerase activities. Mutation of Arg13 into Ala largely reduced the GSH-conjugation activity by approximately 85 - 95%, whereas substitutions by Lys and Leu barely affected activity. These results suggest that, in the GSH-conjugation activity of hGST P1-1, the contribution of Arg13 toward catalytic activity is highly dependent on substrate specificities and the size of the side chain at position 13. From the kinetic parameters, introduction of larger side chains at position 13 results in stronger affinity (Leu > Lys, Arg > Ala) towards GSH. The substitutions of Arg13 with alanine and leucine significantly affected $k_{cat}$, whereas substitution with Lys was similar to that of the wild type, indicating the significance of a positively charged residue at position 13. From the plots of log ($k_{cat}/{K_m}^{CDNB}$) against pH, the $pK_a$ values of the thiol group of GSH bound in R13A, R13K, and R13L were estimated to be 1.8, 1.4, and 1.8 pK units higher than the $pK_a$ value of the wild-type enzyme, demonstrating the contribution of the Arg13 guanidinium group to the electrostatic field in the active site. From these results, we suggest that contribution of Arg13 in substrate binding is highly dependent on the nature of the electrophilic substrates, while in the catalytic mechanism, it stabilizes the GSH thiolate through hydrogen bonding.

Electrochemical Properties of Kaolinite in Aqueous Suspension (수용액중(水溶液中)에서의 Kaolinite 입자(粒子)의 전기화학적(電氣化學的) 성질(性質))

  • Lim, Hyung-Sik;Baham, J.;Volk, V.V.
    • Korean Journal of Soil Science and Fertilizer
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    • v.16 no.4
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    • pp.318-324
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    • 1983
  • Electrochemical properties of Georgia kaolinite in aqueous suspension were studied by ion adsorption, potentiometric titration, and electrophoretic mobility measurements. Kaolinite in 0.001 M and 0.1 M NaCl solution showed qualitatively both pH independent and pH depender negative and positive charges through pH range 2.5-11.0 when dissolved aluminum ions from kaolinite were considered as well as $Na^+$ and $Cl^-$ as index ions. Electrophoretic mobilities (EM) of 0.02 wt. % kaolinite suspension in distilled water and 0.001 M NaCl solution were approximately constant against mobility measuring time consumed in the electrophoresis cell at different pH values, and isoelectric points(IEP) were around pH 4.7. EM values in 0.1 M NaCl solution were positive and constant against mobility measuring time below pH 4; but above pH 4, EM values were negative for the first 10 seconds followed by positive values which became approximately constant through stepped changes after 10 minutes. Hydrated cations may bind to the six- member oxygen ring sites having multiple partial negative charges on the exterior tetrahedral layer surface by both electrostatic and hydrogen bonding force while hydrated anions bind to the partially positively charged hydrogen atoms on the exterior octahedral layer surface. Parts of the aluminol groups on the exterior octahedral layer surface as well as edge faces may be involved in complex reactions and have both anion and cation exchange capacities in the electrolyte solution above pH 4.

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Aminopropyl Functionalized Silica Nanoparticle Dispersed Nafion Composite Membranes for Vanadium Redox Flow Batteries (아미노프로필 관능기를 갖는 실리카 나노 입자가 분산된 나피온 복합막을 이용한 바나듐 레독스 흐름 전지)

  • Lee, Doohee;Yu, Duk Man;Yoon, Sang Jun;Kim, Sangwon;So, Soonyong;Hong, Young Taik
    • Membrane Journal
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    • v.30 no.5
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    • pp.307-318
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    • 2020
  • Conventional perfluorinated sulfonic acid membrane, Nafion is widely used for vanadium redox flow battery (VRFB). It is desired to prevent vanadium ion permeation through a membrane to retain the capacity, and to keep the cell efficiency of a VRFB. Highly proton conductive and chemically stable Nafion membranes, however, suffer from high vanadium permeation, which induce the reduction in charge and discharge capacity by side reactions of vanadium ions. In this study, to resolve the issue, silica nanoparticles, which are functionalized with 3-aminopropyl group (fS) are introduced to enhance the long-term performance of a VRFB by lowering vanadium permeation. It is expected that amine groups on silica nanoparticles are converted to positive ammonium ion, which could deteriorate positively charged vanadium ions' crossover by Gibbs-Donnan effect. There is reduction in proton conductivity may due to acid-base complexation between fS and Nafion side chains, but ion selectivity of proton to vanadium ion is enhanced by introducing fS to Nafion membranes. With the composite membranes of Nafion and fS, VRFBs maintain their discharge capacity up to 80% at a high current density of 150 mA/㎠ during 200 cycles.

Physical properties and intracellular uptake of polyethyleneglycol-incorporated cationic liposomes (폴리에틸렌글리콜이 도입된 양이온성 리포솜의 물리적 특성 및 세포이입효과)

  • Jung, Soon-Hwa;Jung, Suk-Hyun;Kim, Sung-Kyu;Seong, Ha-Soo;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of Pharmaceutical Investigation
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    • v.38 no.1
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    • pp.15-21
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    • 2008
  • Liposomes as one of the efficient drug carriers have some shortcomings such as their short circulation time, fast clearance from human body by reticuloendothelial system (RES) and limited intracellular uptake to target cell. In this study, polyethylenglycol (PEG)-incorporated cationic liposomes were prepared by ionic complexation of positively charged liposomes with carboxylated polyethyleneglycol (mPEG-COOH). The cationic liposomes had approximately $98.6{\pm}1.0nm$ of mean particle diameter and $42.8{\pm}0.8mV$ of zeta potential value. The PEG-incorporated cationic liposomes had $110.1{\pm}1.2nm$ of mean particle diameter with an increase of about 10 nm compared to the cationic liposomes. Zeta potential value of them was $12.9{\pm}0.6mV$ indicating 30mV decrease of cationic charge compared to the cationic liposomes. The amount of PEG which was incorporated onto the cationic liposomes was assayed by using picrate assay method and the incorporation efficiency was $58.4{\pm}1.1%$. Loading efficiency of model drug, doxorubicin, into cationic liposomes or PEG-incorporated cationic liposomes was about $96.0{\pm}0.7%$. Results of intracellular uptake which were evaluated by flow cytometry analysis of doxorubicin loaded liposomes showed that intracellular uptake of PEG-incorporated cationic liposomes was higher than the cationic liposomes or DSPE-mPEG liposomes. In addition, cytotoxicity of PEG-incorporated cationic liposomes was comparable to cationic liposomes. Consequently, the PEG-incorporated cationic liposomes of which surface was incorporated with PEG by ionic complex may be applicable as anticancer drug carriers that can increase therapeutic efficacy.

A Kinetics Study of Rn Daughter and Atmospheric Trace Gas Using Alpha Track Detection (알파비적검출방법에 의한 대기중 라돈딸핵종의 화학적 동특성연구)

  • Yoon, Suk-Chul;Ha, Chung-Woo
    • Journal of Radiation Protection and Research
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    • v.20 no.2
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    • pp.79-83
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    • 1995
  • A number of investigators have reported formation of radiolytic ultrafine particles produced by the interaction of ionizing radiation with atmospheric trace gases. Previous studies have suggested that a very high localized concentration of the hydroxyl radical produced by the radiolysis of water can react with atmospheric trace gases such as $SO_2$ and produce lower vapor pressure compounds that can subsequently nucleate. To determine the trace gas and water vapor concentration dependence of the active, positively charged, first decayt product of radon (Po-218), a well-controlled radon chamber was used in this research. The mobility spectrum of the decay products in the range of $0.07-5.0cm^2/V\;sec$ from the radon chamber was measured using alpha track detector installed inside a specially-designed electrostatic spectrometer. Measurements were taken for different concentrations (0.5ppm to 5ppm) of $SO_2$ in Purified, Compressed air. A kinetics Study following the clustering of $SO_2$ around the $PoO_x^+$ ion in an excess of $SO_2$ for interpretation of the reaction processes was performed.

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Characterization of immobilized laccase and its catalytic activities (고정된 laccase의 특성 및 촉매효과)

  • Hyung Kyung Hee;Shin Woonsup
    • Journal of the Korean Electrochemical Society
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    • v.2 no.1
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    • pp.31-37
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    • 1999
  • Copper-containig enzyme, laccase (Rhus vernicifera) was immobilized onto gold electrode using self-assembly technique and its surface properties and catalytic activities were examined. Laccase is an oxidoreductase capable to oxidize diphenols or diamines by 4-electron reduction of molecular oxygen without superoxide or peroxide intermediates. The electrode surface were modified by $\beta-mercaptopropionate$ to have a net negative charge in neutral solution and positively charged laccase (pI=9) was immobilized by electrostatic interaction. The successful immobilization was confirmed by cyclic voltammograms which showed typical surface-confined shapes and behaviors. The amount of charge to reduce the surface was similar to the charge calculated assuming the surface being covered by monolayer. The activity of the immobilized enzyme was tested by the capbility of oxidizing a substrate, ABTS (2,2-azine-bis-(3-ethylbenzthioline-6-sulfonic acid) and it was maintained for $2\~3$ days at $4^{\circ}C$. The immobilzed laccase showed about $10\~15\%$ activity compared to that in solution. The laccase-modified electrode showed the activity of elefoocatalytic reduction of oxygen in the presence of mediator, $Fe(CN)_6^{3-}$ The addtion of azide which is an inhibitor of laccase compeletly eliminated the catalytic current.