• 제목/요약/키워드: electron paramagnetic resonance

검색결과 96건 처리시간 0.023초

Interaction of Cytochrome c and Cytochrome c Oxidase Studied by Spin-Label EPR and Site-Directed Mutagenesis

  • Park, Hee-Young;Chun, Sun-Bum;Han, Sang-Hwa;Lee, Kwang-Soon;Kim, Kyung-Hoon
    • BMB Reports
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    • 제30권6호
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    • pp.397-402
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    • 1997
  • A thiol-specific spin label was attached to cysteine-102 of yeast cytochrome c and electron paramagnetic resonance (EPR) spectra were measured as a function of added cytochrome c oxidase concentration. The intensity decreased due to line broadening as cytochrome c formed a complex with cytochrome c oxidase and reached a minimum when the ratio of cytochrome c to cytochrome c oxidase became one. Replacement of either Lys-72 or Lys-87 of cytochrome c by Glu did not result in a significant change in binding affinity. Interestingly the K72E mutant, unlike K87E, had a much lower rate of electron transfer than the wild type. These results indicate that many positively charged residues as a group participate in complex formation but Lys-72 might be important for cytochrome c to be locked in an orientation for an efficient electron transfer. A stoichiometry of 1 was also confirmed by optical absorption of the cytochrome c-cytochrome c oxidase complex which had been run through a gel chromatography cloumn to remove unbound cytochrome c. The EPR spectrum of this 1:1 complex, however, was a mixture of two components. This explains a biphasic kinetics for a single binding site on cytochrome c oxidase without invoking conformational transition.

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The Effect of Gamma Irradiation on PLGA and Release Behavior of BCNU from PLGA Wafer

  • Lee, Jin-Soo;Chae, Gang-Soo;Gilson Khang;Kim, Moon-Suk;Cho, Sun-Hang;Lee, Hai-Bang
    • Macromolecular Research
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    • 제11권5호
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    • pp.352-356
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    • 2003
  • The objectives of this study were to investigate the influence of gamma irradiation for sterilization on poly(D,L-lactide-co-glycolide) (PLGA) with different molecular weight and the effect of gamma irradiation on the release behavior of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, carmustine) from PLGA wafer with various irradiation doses. The effect of gamma irradiation on PLGA was evaluated by gel permeation chromatography (GPC), differential scanning calorimetry (DSC), and electron paramagnetic resonance (EPR). The weight average molecular weight (M$_{w}$) and glass transition temperature (T$_{g}$) of PLGA decreased after gamma irradiation. The extent of M$_{w}$ reduction was dependent on irradiation dose and PLGA molecular weight. Using EPR spectroscopy, we successfully detected gamma irradiation induced free radicals in PLGA. The gamma irradiation increased the release rate of BCNU from PLGA wafer at applied irradiation doses except 2.5 Mrad of irradiation dose in this study.study.

High Pressure Synthesis and Physical Properties of the Solid Solution, $SrLaAl_{1-x}Ni_xO_4(0

  • 변송호
    • Bulletin of the Korean Chemical Society
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    • 제16권11호
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    • pp.1084-1088
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    • 1995
  • A complete solid solution (SrLaAl1-xNixO4) between insulating SrLaAlO4 and metallic SrLaNi(Ⅲ)O4 oxides were prepared under high oxygen pressure (1.5 kbar, 800 ℃). They have tetragonal K2NiF4-type structure in all the solid solution range. Compared with lattice parameters of the same solid solution prepared under normal condition (1 bar, 1200 ℃), large decrease in the c-parameter was induced by high pressure treatment while no noticeable variation of the a-parameter was observed. Although marked changes of structural parameters, magnetic susceptibilities, and electron paramagnetic resonance spectra were consistently occurred before and after x=0.5, overall behaviors were essentially the same with those of solid solution prepared under normal condition. Such a phenomenon is explained by assuming the formation of partially filled narrow σ*x2-y2 band for x>0.5. Lattice contraction along the c-axis by high pressure treatment seems not to broaden this band. Particularly, the continuous absorption characteristic of a high free carrier concentration for x>0.5 and the absence of Ni-O in-plane stretching mode in the infrared absorption spectra supports this picture. However, the conductivities increasing with temperature for all solid solution suggest that some localization character, of probably Anderson type, remains for x>0.5.

PbWO4 : Gd 단결정 내의 Gd3+ 상자성 이온에 대한 바닥 상태 에너지 (Ground State Energy of Gd3+ Paramagnetic Ion in PbWO4 : Gd Single Crystal)

  • 염태호
    • 한국자기학회지
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    • 제26권2호
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    • pp.45-49
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    • 2016
  • 분광학적 분리인자 $g_{ij}$와 영자기장 갈라지기 상수 값 $B_k^q$ 값을 유효 스핀 하밀토니안에 사용하여, $PbWO_4$ : Gd 단결정 내의 정방정계 대칭성 자리에 위치하고 있는 $Gd^{3+}$ 상자성 불순물 이온(유효전자 스핀 S = 7/2)의 바닥상태에서의 에너지 준위를 계산하였다. 외부 자기장이 영일 경우의 $Gd^{3+}$ 이온의 영자기장 갈라지기 값은 $PbWO_4$ : Gd 단결정의 방향에 관계없이 모두 같았고, 이때 ${\mid{\pm}7/2}$ > ${\leftrightarrow}{\mid{\pm}5/2}$ >, ${\mid{\pm}5/2}$ > ${\leftrightarrow}{\mid{\pm}3/2}$ >, ${\mid{\pm}3/2}$ > ${\leftrightarrow}{\mid{\pm}1/2}$ > 전이 사이에서 계산된 에너지 간격은 각각, 6.9574 GHz, 6.9219 GHz, 15.8704 GHz이다. 결정학적 축에 대하여 외부 자기장을 가하는 방향에 따라서 서로 다른 에너지 준위 값을 나타내었다. 이중 외부자기장이 결정학적 주축 a- 및 c-축에 나란할 경우에 에너지 준위를 계산하여 논의하였다.

Sol-Gel Synthesis, Crystal Structure, Magnetic and Optical Properties in ZnCo2O3 Oxide

  • Das, Bidhu Bhusan;Barman, Bittesh
    • 대한화학회지
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    • 제63권6호
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    • pp.453-458
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    • 2019
  • Synthesis of ZnCo2O3 oxide is performed by sol-gel method via nitrate-citrate route. Powder X-ray diffraction (XRD) study shows monoclinic unit cell having lattice parameters: a = 5.721(1) Å, b = 8.073(2) Å, c = 5.670(1) Å, β = 93.221(8)°, space group P2/m and Z = 4. Average crystallite sizes determined by Scherrer equation are the range ~14-32 nm, whereas SEM micrographs show nano-micro meter size particles formed in ZnCo2O3. Endothermic peak at ~798 K in the Differential scanning calorimetric (DSC) trace without weight loss could be due to structural transformation and the endothermic peak ~1143 K with weight loss is due to reversible loss of O2 in air atmosphere. Energy Dispersive X-ray (EDX) analysis profile shows the presence of elements Zn, Co and O which indicates the purity of the sample. Magnetic measurements in the range of +12 kOe to -12 kOe at 10 K, 77 K, 120 K and at 300 K by PPMS-II Physical Property Measurement System (PPMS) shows hysteresis loops having very low values of the coercivity and retentivity which indicates the weakly ferromagnetic nature of the oxide. Observed X-band EPR isotropic lineshapes at 300 K and 77 K show positive g-shift at giso ~2.230 and giso ~2.217, respectively which is in agreement with the presence of paramagnetic site Co2+(3d7) in the oxide. DC conductivity value of 2.875 ×10-8 S/cm indicates very weakly semiconducting nature of ZnCo2O3 at 300 K. DRS absorption bands ~357 nm, ~572 nm, ~619 nm and ~654 nm are due to the d-d transitions 4T1g(4F)→2Eg(2G), 4T1g(4F)→4T1g(4P), 4T1g(4F)→4A2g(4F), 4T1g(4F)→4T2g(4F), respectively in octahedral ligand field around Co2+ ions. Direct band gap energy, Eg~ 1.5 eV in the oxide is obtained by extrapolating the linear part of the Tauc plot to the energy axis indicates fairly strong semiconducting nature of ZnCo2O3.

Irradiation Detection in Korean Traditional Soybean-Based Fermented Powdered Sauces: Data for Establishing a Database for Regulation of Irradiated Foods

  • Choi, In-Duck;Kim, Byeong-Keun;Song, Hyun-Pa;Byun, Myung-Woo;Kim, Myung-Cheol;Lee, Jong-Oak;Lee, Ho-Joon;Kim, Dong-Ho
    • Preventive Nutrition and Food Science
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    • 제10권1호
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    • pp.29-33
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    • 2005
  • To facilitate establishing regulations for irradiated foods, Korean traditional soybean-based fermented powdered doenjang (PD), kanjang (PK), kochujang (PKC) and chungkukjang (PC) were irradiated at 1, 3, 5 and 7 kGy, and subjected to irradiation detection analyses as part of establishing a database for detecting irradiated foods. Photostimulated luminescence (PSL) and electron spin resonance (ESR) were applied as the detection methods. Using PSL analysis, the irradiated PD, PK and PKC could be easily distinguished from the non-irradiated ones, while irradiation of the PC at 5 kGy or higher was detectable. The ESR spectra of the irradiated PD, PK and PKC exhibited symmetrical multiplet lines, which might be induced from the crystalline sugar, whereas, the PC showed a single signal at the paramagnetic centers. The signal intensity increased with incremental increases of irradiation doses distinguishing the irradiated samples from the control. In addition, the peak height also revealed that irradiation induced an increment in the intensity of single and/or multiplet lines of the ESR signals, resulting in clear confirmation of irradiation. Thus, the data from this study could be used as references for detecting irradiated soybean-based fermented powdered sauces.

감마선 조사된 남양진주의 검지 (Detection of gamma irradiated South Sea cultured pearls)

  • 최현민;이보현;김영출
    • 한국결정성장학회지
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    • 제22권1호
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    • pp.36-41
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    • 2012
  • 남양진주에 $Co^{60}$을 이용하여 0.1~100 kGy까지 선량의 범위에서 방사선 조사 실험을 행하였다. 더불어 방사선 조사된 남양진주의 검지를 위해 전자상자성공명(EPR)과 아미노산분석(AAA)을 행하였다. EPR 분석에서 방사선 조사후, 방사선 조사전에는 없었던 free radical이 생성되었으며 $CO_2^-$ radical의 g-factor는 $2.001{\pm}0.002$이었다. 진주층의 아미노산 분석 결과 방사선 조사후의 glutamic acid는 11.43 %, alanine은 3.11 %, histidine은 43.75 %의 감소량을 보였으며, 이는 일부 아미노산이 파괴되었음을 보여준다. 본 연구의 결과로 EPR 분석은 방사선 조사된 남양진주의 검지에 적합하였다.

Radical Intermediate Generation and Cell Cycle Arrest by an Aqueous Extract of Thunbergia Laurifolia Linn. in Human Breast Cancer Cells

  • Jetawattana, Suwimol;Boonsirichai, Kanokporn;Charoen, Savapong;Martin, Sean M
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권10호
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    • pp.4357-4361
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    • 2015
  • Thunbergia Laurifolia Linn. (TL) is one of the most familiar plants in Thai traditional medicine that is used to treat various conditions, including cancer. However, the antitumor activity of TL or its constituents has never been reported at the molecular level to support the folklore claim. The present study was designed to investigate the antitumor effect of an aqueous extract of TL in human breast cancer cells and the possible mechanism(s) of action. An aqueous crude extract was prepared from dried leaves of TL. Folin-Ciocalteu colorimetric assays were used to determine the total phenolic content. Antiproliferative and cell cycle effects were evaluated in human breast adenocarcinoma MCF-7 cells by MTT reduction assay, cell growth inhibition, clonogenic cell survival, and flow cytometric analysis. Free radical generation by the extracts was detected using electron paramagnetic resonance spectroscopy. The exposure of human breast adenocarcinoma MCF-7 cells to a TL aqueous extract resulted in decreases in cell growth, clonogenic cell survival, and cell viability in a concentration-dependent manner with an $IC_{50}$ value of $843{\mu}g/ml$. Treatments with extract for 24h at $250{\mu}g/ml$ or higher induced cell cycle arrest as indicated by a significant increase of cell population in the G1 phase and a significant decrease in the S phase of the cell cycle. The capability of the aqueous extract to generate radical intermediates was observed at both high pH and near-neutral pH conditions. The findings suggest the antitumor bioactivities of TL against selected breast cancer cells may be due to induction of a G1 cell cycle arrest. Cytotoxicity and cell cycle perturbation that are associated with a high concentration of the extract could be in part explained by the total phenolic contents in the extract and the capacity to generate radical intermediates to modulate cellular proliferative signals.

Membrane Topology of Helix 0 of the Epsin N-terminal Homology Domain

  • Kweon, Dae-Hyuk;Shin, Yeon-Kyun;Shin, Jae Yoon;Lee, Jong-Hwa;Lee, Jung-Bok;Seo, Jin-Ho;Kim, Yong Sung
    • Molecules and Cells
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    • 제21권3호
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    • pp.428-435
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    • 2006
  • Specific interaction of the epsin N-terminal homology(ENTH) domain with the plasma membrane appears to bridge other related proteins to the specific regions of the membrane that are invaginated to form endocytic vesicles. An additional $\alpha$-helix, referred to as helix 0 (H0), is formed in the presence of the soluble ligand inositol-1,4,5-trisphosphate [$Ins(1,4,5)P_3$] at the N terminus of the ENTH domain (amino acid residues 3-15). The ENTH domain alone and full-length epsin cause tubulation of liposomes made of brain lipids. Thus, it is believed that H0 is membrane-inserted when it is coordinated with the phospholipid phosphatidylinositol-4,5-bisphosphate [$PtdIns(4,5)P_2$], resulting in membrane deformation as well as recruitment of accessory factors to the membrane. However, formation of H0 in a real biological membrane has not been demonstrated. In the present study, the membrane structure of H0 was determined by measurement of electron paramagnetic resonance (EPR) nitroxide accessibility. H0 was located at the phosphate head-group region of the membrane. Moreover, EPR line-shape analysis indicated that no pre-formed H0-like structure were present on normal acidic membranes. $PtdIns(4,5)P_2$ was necessary and sufficient for interaction of the H0 region with the membrane. H0 was stable only in the membrane. In conclusion, the H0 region of the ENTH domain has an intrinsic ability to form H0 in a $PtdIns(4,5)P_2$-containing membrane, perhaps functioning as a sensor of membrane patches enriched with $PtdIns(4,5)P_2$ that will initiate curvature to form endocytic vesicles.

EPR Studies of the Active Sites of Carbon Monoxide Dehydrogenase from Clostridium thermoaceticum

  • Shin, Woonsup;Lindahl, Paul A.
    • 분석과학
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    • 제8권4호
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    • pp.869-876
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    • 1995
  • The active sites of the nickel and iron-containing enzyme, carbon monoxide dehydrogenase (CODH) from clostridium thermoaceticum were investigated using Electron Paramagnetic Resonance (EPR) technique. CODH exhibits several spectral features called NiFeC, $g_{ave}=1.82$, $g_{ave}=1.86$. FCII signals which are originated from different clusters in this enzyme. CODH is know to catalyze two different kinds of reactions - acetyl-CoA synthesis and CO oxidation. The acetyl-CoA synthesis activity can be followed by monitoring CO/acetyl-CoA exchange. The addition of 1,10-phenanthroline (phen) to CODH selectively destroyed the CO/acetyl-CoA exchange activity and eliminated the NiFeC signal completely. CO oxidation activity and other EPR signals were unaffected. Such behavior demonstrates that CODH has two distinct active sites and that the NiFe complex is only responsible for the CO/acctyl-CoA exchange activity. Phen caused the removal of only 30% of Ni in the NiFe complex ($0.3Ni/{\alpha}{\beta}$) as shown by the quantitative metal analysis. The phen-treated CODH could be reactivated fully by incubation In $Ni^{2+}$ solution. Radioactive $^{63}Ni^{2+}$ was used to quantitate the amount of the $Ni^{2+}$ incorporated into phen-treated enzyme and showed that the amount was the same as the removed by the phen treatment. i.e. $0.3Ni/{\alpha}{\beta}$. This indicates that only 30% of NiFe complexes are labile and responsible for the CO/acctyl-CoA exchange activity, the other 70% are non-labile and have no exchange activity. This is the first clear evidence that the NiFe complex is heterogencous and labile and non-labile Ni sites arc interacting differently with substrates and chelating agents like phen.

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