• Title/Summary/Keyword: Electrostatic field

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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.

Development of Dispenser System with Electrohydrodynamic and Voice Coil Motor for White Light Emitting Diode (백색 LED 제조를 위한 정전기력과 보이스코일모터를 이용한 디스펜서 시스템 개발)

  • Kang, Dong-Seong;Kim, Ki-Beom;Ha, Seok-Jae;Cho, Myeong-Woo;Lee, Woo-Jung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.10
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    • pp.6925-6931
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    • 2015
  • LED(Light Emitting Diode) is used in various filed like a display because of low power consuming, long life span, high brightness, rapid response time and environmental-friendly characteristic. General fabrication method is combination blue light LED chip with yellow fluorescent substance. Because this way is suitable for industry field in terms of convenience, economic, efficiency. In white light LED packaging process, encapsulation process that is dispensing fluorescent substance with silicon to blue light LED chip is most important. So, in this paper we develop EHD pump system using voice coil motor and electrostatic pump for dispensing fluorescent substance. For these things we conduct basic test about liquid surface profiles by voltage and process time. Through this data we decide optimal process condition and verify the optimal condition using design of experiment method. And to confirm uniformity of the condition, we conduct repeat dispensing test.

Preparation and Characterization of Reduced Graphene Oxide with Carboxyl Groups-Gold Nanorod Nanocomposite with Improved Photothermal Effect (향상된 광열 효과를 갖는 카르복실화된 환원 그래핀옥사이드-골드나노막대 나노복합체의 제조 및 특성 분석)

  • Lee, Seunghwa;Kim, So Yeon
    • Applied Chemistry for Engineering
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    • v.32 no.3
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    • pp.312-319
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    • 2021
  • Photothermal therapy is a treatment that necrotizes selectively the abnormal cells, in particular cancer cells, which are more vulnerable to heat than normal cells, using the heat generated when irradiating light. In this study, we synthesized a reduced graphene oxide with carboxyl groups (CRGO)-gold nanorod (AuNR) nanocomposite for photothermal treatment. Graphene oxide (GO) was selectively reduced and exfoliated at high temperature to synthesize CRGO, and the length of AuNR was adjusted according to the amount of AgNO3, to synthesize AuNR with a strong absorption peak at 880 nm, as an ideal photothermal agent. It was determined through FT-IR, thermogravimetric and fluorescence analyses that more carboxyl groups were conjugated with CRGO over RGO. In addition, CRGO exhibited excellent stability in aqueous solutions compared to RGO due to the presence of carboxylic acid. The CRGO-AuNR nanocomposites fabricated by electrostatic interaction have an average size of ~317 nm with a narrow size distribution. It was confirmed that under radiation with a near-infrared 880 nm laser which has an excellent tissue transmittance, the photothermal effect of CRGO-AuNR nanocomposites was greater than that of AuNR due to the synergistic effect of the two photothermal agents, CRGO and AuNR. Furthermore, the results of cancer cell toxicity by photothermal effect revealed that CRGO-AuNR nanocomposites showed superb cytotoxic properties. Therefore, the CRGO-AuNR nanocomposites are expected to be applied to the field of anticancer photothermal therapy based on their stable dispersibility and improved photothermal effect.