• Title/Summary/Keyword: 금속결합 단백질

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Polarographic behavior of Cd(II) and Ni(II) ions in the presence of imidazole groups(II) (Imidazole 및 그유도체의 금속착염에 관한 Polarograph 연구 (II))

  • Kim, Joon Yong;Shin, Joung Hee
    • YAKHAK HOEJI
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    • v.16 no.2
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    • pp.73-84
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    • 1972
  • 단백질과 전이금속 이온들과의 상호작용을 연구하는데 있어서 단백질중의 이미다졸기 및 아미노산의 반응성은 중요하다. Cd(II)-2-메틸이미다졸 착물들의 경우 이미다졸 분자중의 결합측은 "피로올"형 보다 오히려 "피리딘" 질소임을 알 수 있다. 이 착물들의 환원파는 가역적이며 2전자반응으로 나타난다. 그리고 이미다졸 존재하의 Ni(II) 이온은 촉매파를 나타내며 Ni(II) 이온만의 환원파는 불가역성인 것에 비해 가역성을 보여주었다. 또한 그 반파전위는 Ni(II) 이온만의 반파전위보다 수백 mV 양전위로 이동된다. 이러한 성질은 전극 반응에 관여한 주물질이 전극 표면에서 흡착되어 환원되기 때문이다. 이미다졸 농도의 대수값에 대해 반파전위를 도시했을 때 그 기울기가 30mV로써 이론치와 일치한다. 한편 $Cd(II)-2CH_{3}Im$의 단계적 생성항수 $pK_{1},pK_{2},pK_{3},pK_{4}$는 각각 2.68, 4.01, 4.90, 5.36이었다.4.01, 4.90, 5.36이었다.

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Effect of Trace Metal on Accumulation and Physiological Response of the Polychaete, Perinereis nuntia (미량금속 노출에 따른 갯지렁이(Perinereis nuntia)의 미량금속 축적 및 생리학적인 반응)

  • Won, Eun-Ji;Ra, Kong-Tae;Hong, Seong-Jin;Kim, Kyung-Tae;Lee, Jae-Seong;Shin, Kyung-Hoon
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.13 no.4
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    • pp.288-295
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    • 2010
  • Metal exposure experiments using polychaete (Perinereis nuntia) as a bio-indicator of trace metals contamination were conducted to evaluate the bioaccumulation and the biomarkers responses such as metallothionein-like protein (MTLPs) and glutathione S-transferase (GST) which was simultaneously exposed to Cadmium (Cd) and Copper (Cu). Cu and Cd concentrations in polychaete were enhanced with increasing exposure time and their concentrations of aqueous medium. Initial accumulation of Cd was higher than that of Cu. Our results showed that the bioaccumulation of Cu and Cd were prohibited, especially at higher Cu levels, suggesting the different cellular uptake mechanisms when Cu and Cd are co-exist. Net accumulation rate of Cu was declined with exposure time but it did not show any significant change for Cd. Although the highest MTLPs concentration was observed at 6 hr of exposure time, it did not show any significant change related to exposure times and metals concentrations. An increase of GST activity tended to increase as a function of exposure time and metals concentrations. And GST activities in P. nuntia have similar tendency with bioconcentration factors in high concentration of Cu (treatment group IV) at post 24 h of exposure. Our results provide new information of the bioaccumulation and biomarker responses to understand the effects of co-existing contaminants (Cu and Cd) using polychaete. Further studies are required to elucidate the bioaccumulation and biomarkers responses for various contaminants.

Heterocyclic Amines Removal by Binding Ability of Lactic Acid Bacteria Isolated from Soybean Paste (된장에서 분리된 유산균의 결합력에 의한 Heterocyclic Amines 제거)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.73-83
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    • 2014
  • The objective of the this study was to investigate the binding capacity and removal ability of lactic acid bacterial strains obtained from Korean soybean paste for mutagenic heterocyclic amines (HCAs) formed during cooking of protein-rich food at high temperature. Among 19 strains identified by carbohydrate fermentation and 16S rRNA sequencing, the live cell or cell-free culture supernatant of Lactobacillus acidophilus D11, Enterococcus faecium D12, Pediococcus acidilactici D19, L. acidophilus D38, Lactobacillus sakei D44, Enterococcus faecalis D66, and Lactobacillus plantarum D70 inhibited the mutagenesis caused by either 3-amino-1,4-dimethyl-5H-pyrido[4,3-b] indole (Trp-P-1) or 3-amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) in Salmonella typhimurium TA98 and TA100. The bacterial cells of the isolated strains showed greater binding activity than the pure cell wall, exopolysaccharide, and pepetidoglycan. The carbohydrate moieties of the cell wall or protein molecules on the cell surface have a significant role in binding Trp-P-1 and Trp-P-2, since protease, heating, sodium metaperiodate, or acidic pH treatments significantly (P<0.05) reduced the binding efficacy of the tested bacteria. Addition of metal ions or sodium dodecyl sulfate decreased the binding ability of E. faecium D12, L. acidophilus D38, and E. faecalis D66. Therefore, the binding mechanisms of these strains may consist of ion-exchange and hydrophobic bonds. Especially, the high mutagen binding by L. acidophilus D38 and L. plantarum D70 may reduce the accumulation or absorption of Trp-P-1 and Trp-P-2 in the small intestine via increased excretion of a mutagen-bacteria complex.

Phosphoprotein Partitioning in Metal-Affinity Aqueous Two-Phase Systems and Prediction of Partitioning Behavior (금속 친화성 액 이상분계 시스템에서 Phsphoprotein분배 및 분배예측)

  • 정봉현
    • KSBB Journal
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    • v.9 no.3
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    • pp.279-286
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    • 1994
  • A mathematical model has been derived and used to describe phosphoprotein partitioning in Fe(III) IDA-PEG/dextran two-phase systems. This model includes the inhibitory effects of hydrogen and hydroxyl ion concentrations on protein partitioning. For aqueous two-phase partitioning experiments, the Al and A2 subcomponents of ovalbumin carrying two and one surface phosphoryl group(s) were purified using an immobilized metal ion affinity chromatography (IMAC). The ratio of partition coefficients in the presence and absence of Fe(III)IDA-PEG, K/Ko, increased in the pH range of 3.0 to 5.0 due to deprotonation of the second oxygen of the phosphoryl group, and above pH 5.0 declined steeply by the inhibitory binding of hydroxyl ions to the metal ion. This partitioning behavior was well described by the mathematical model. The binding constants for formation of the complex between the phosphoryl group and the Fe(III)IDA-PEG were found to be $6.1{\times}10^3M^{-1} and 2.3{\times}10^4M^{-1}$ in the top and bottom phases, respectively. These values are 3-5 times those for interaction of Cu(II)IDA-PEG with a single surface-accessible histidine.

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Protein structure analysis : Pharmacophore study for new insecticide target AnCE using the substrate of ACE, HHL molecule (단백질의 구조연구 : ACE의 기질 HHL을 이용한 신규 살충제 표적 AnCE에 대한 약리단 연구)

  • Lee, Jung-Kyung;Kim, Kyeong-Yee
    • The Korean Journal of Pesticide Science
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    • v.9 no.3
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    • pp.191-198
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    • 2005
  • Hippuryl-L-histidyl-L-leucine (Hip-L-His-L-Leu, HHL) is the known substrate of ACE, which used often in inhibition kinetic study to design new inhibitor. Here we use HHL molecule as a template to predict pharmacophore which can interact with residues in active site of AnCE, new potential insecticide target protein. To explain physicochemical properties related to molecular geometry and conformational change in reaction field as well as electron density of atoms associated to pharmacophores, geometry optimization, NMR chemical shifts and natural population analysis were performed by ab initio and DFT method. Calculated NMR chemical shifts showed good agreement with the experimental ones and obtained electron densities were used for analyzing pharmacophores of corresponding atoms. Finally, we could extract aye pharmacophores related to hydrophobic aliphatic and aromatic site, hydrogen bonding donor and acceptor site and Zn binding site from the HHL molecule.

Radiolabeling of NOTA and DOTA with Positron Emitting $^{68}$Ga and Investigation of In Vitro Properties (양전자 방출핵종 $^{68}$Ga을 이용한 NOTA와 DOTA의 표지 및 시험관내 특성 연구)

  • Jeong, Jae-Min;Kim, Young-Ju;Lee, Yun-Sang;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.330-336
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    • 2009
  • Purpose: We established radiolabeling conditions of NOTA and DOTA with a generator-produced PET radionuclide $^{68}$Ga and studied in vitro characteristics such as stability, serum protein binding, octanol/water distribution, and interference with other metal ions. Materials and Methods: Various concentrations of NOTA 3HCl and DOTA 4HCl were labeled with 1 mL $^{68}$GaCl$_3$ (0.18$\sim$5.75 mCi in 0.1 M HCl in various pH. NOTA 3HCl (0.373 mM) was labeled with $^{68}$GaCl$_3$(0.183$\sim$0.232 mCi/0.1 M HCl 1.0 mL) in the presence of CuCl$_2$, FeCl$_2$, InCl$_3$, FeCl$_3$, GaCl$_3$, MgCl$_2$ or CaCl$_2$ (0$\sim$6.07 mM) at room temperature. The labeling efficiencies of $^{68}$Ga-NOTA and $^{68}$Ga-DOTA were checked by ITLC-SG using acetone or saline as mobile phase. Stabilities, protein bindings, and octanol distribution coefficients of the labeled compounds also were investigated. Results: $^{68}$Ga-NOTA and $^{68}$Ga-DOTA were labeled optimally at pH 6.5 and pH 3.5, respectively, and the chelates were stable for 4 hr either in the reaction mixture at room temperature or in the human serum at 37$^{\circ}C$. NOTA was labeled at room temperature while DOTA required heating for labeling. $^{68}$Ga-NOTA labeling efficiency was reduced by CuCl$_2$, FeCl$_2$, InCl$_2$, FeCl$_3$ or CaCl$_3$, however, was not influenced by MgCl$_2$ or CaCl$_2$. The protein binding was low (2.04$\sim$3.32%). Log P value of $^{68}$Ga-NOTA was -3.07 indicating high hydrophilicity. Conclusion: We found that NOTA is a better bifunctional chelating agent than DOTA for $^{68}$Ga labeling. Although, $^{68}$Ga-NOTA labeling is interfered by various metal ions, it shows high stability and low serum protein binding.

Zero-Length Crosslinking Study on Interactions of TBP, GAL4-AH, and TFIIB in the Preinitiation Complex (전사개시전 복합체에서 TBP, GAL4-AH, TFIIB의 상호작용에 대한 Zero-Length Crosslinking 실험)

  • 권혁만
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.393-399
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    • 1996
  • A zero-Iength croessinking procedure for studying protein-protein interactlons in preinitiation complex has heen developed. Preinltiadon complexes were assembled with immobilized DNA templates coupled to metal beads. Pudfied complexes were dIrectly crosslinked by 1-ethyl-3.(3-dimethylaminopropyl)carbodlimide (EDC). The reaction was stopped by addition of $\beta$-mercaptoethanol, and the complexes were isolated from EDC immedIately. An appllcatlion of this method with a preinltlation complex assembled with TBP, TFIIB, and GAL4-AH demonstrated that ThP dIrectly interacts with GAL4-AH and TFIIB in the prelnitlatlon complex. However, croeslinked produd between GAL4-AH and ThilB was not observed. These resutts lndlcate that GAL4-AH does not stably Interact with TFIIB In the GAL4-AH-TFIIB-TBP-DNA prelnitlatlon complex.

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Characterization of extracellular protease from Pseudoxanthomonas sp. WD12 and WD32 (Pseudoxanthomonas sp. WD12와 WD32의 세포외 단백질분해효소 특성)

  • Cho, Woon-Dong;Oh, Ji-Sung;Roh, Dong-Hyun
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.285-288
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    • 2016
  • Proteolytic enzymes perform hydrolysis of the peptide bonds in the protein and most commonly use in the industry. Pseudoxanthomonas sp. WD12 and WD32 were previously isolated as protease producers from a rotten wood sample. Here, we report the secreted proteolytic enzymes. The optimum enzyme reaction temperature for the secreted crude enzyme from the strain WD12 and WD32 were $50^{\circ}C$ at pH 9.0 and $45^{\circ}C$ at pH 8.0, respectively. The enzyme activities of both strains were increased by addition of KCl, NaCl, $CaCl_2$ or $MnSO_4$, and decreased by addition of $AgNO_3$, $CuSO_4$, $FeCl_3$ or $AlCl_3$. Secreted enzymes of both strains were most strongly inhibited by addition of $FeCl_3$ or $CuSO_4$. Taken together these results, WD12 could be a candidate strain of industrial alkaline protease production.

다채널 표면 플라즈몬 공명 영상장치를 이용한 자기조립 단분자막의 표면 분석

  • Pyo, Hyeon-Bong;Sin, Yong-Beom;Yun, Hyeon-Cheol
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.74-78
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    • 2003
  • Multi-channel images of 11-MUA and 11-MUOH self-assembled monolayers were obtained by using two-dimensional surface plasmon resonance (SPR) absorption. Patterning process was simplified by exploiting direct photo-oxidation of thiol bonding (photolysis) instead of conventional photolithography. Sharper images were resolved by using a white light source in combination with a narrow bandpass filter in the visible region, minimizing the diffraction patterns on the images. The line profile calibration of the image contrast caused by different resonance conditions at each points on the sensor surface (at a fixed incident angle) enables us to discriminate the monolayer thickness in sub-nanometer scale. Furthermore, there is no signal degradation such as photo bleaching or quenching which are common in the detection methods based on the fluorescence.

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Organic Memory Device Using Self-Assembled Monolayer of Nanoparticles (나노입자 자기조립 단일층을 이용한 유기메모리 소자)

  • Jung, Hunsang;Oh, Sewook;Kim, Yejin;Kim, Minkeun;Lee, Hyun Ho
    • Applied Chemistry for Engineering
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    • v.23 no.6
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    • pp.515-520
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    • 2012
  • In this review, the fabrication of silicon based memory capacitor and organic memory thin film transistors (TFTs) was discussed for their potential identification tag applications and biosensor applications. Metal or non-metal nanoparticles (NPs) could be capped with chemicals or biomolecules such as protein and oligo-DNA, and also be self-assembly monolayered on corresponding target biomolecules conjugated dielectric layers. The monolayered NPs were formed to be charging elements of a nano floating gate layer as forming organic memody deivces. In particular, the strong and selective binding events of the NPs through biomolecular interactions exhibited effective electrostatic phenomena in memory capacitors and TFTs formats. In addition, memory devices fabricated as organic thin film transistors (OTFTs) have been intensively introduced to facilitate organic electronics era on flexible substrates. The memory OTFTs could be applicable eventually to the development of new conceptual devices.