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Analytical biochemistry 13건

  1. [해외논문]   New method for estimating clustering of DNA lesions induced by physical/chemical mutagens using fluorescence anisotropy   SCI SCIE

    Akamatsu, Ken (Radiation DNA Damage Research Group, Kansai Photon Science Institute, National Institutes for Quantum and Radiological Science and Technology (QST), 8-1-7 Umemidai, Kizugawa, Kyoto 619-0215, Japan ) , Shikazono, Naoya (Radiation DNA Damage Research Group, Kansai Photon Science Institute, National Institutes for Quantum and Radiological Science and Technology (QST), 8-1-7 Umemidai, Kizugawa, Kyoto 619-0215, Japan ) , Saito, Takeshi (Radiation Biochemistry and Biological Function, Research Reactor Institute, Kyoto University, Kumatori, Sennan, Osaka 590-0494, Japan)
    Analytical biochemistry v.536 ,pp. 78 - 89 , 2017 , 0003-2697 ,

    초록

    Abstract We have developed a new method for estimating the localization of DNA damage such as apurinic/apyrimidinic sites (APs) on DNA using fluorescence anisotropy. This method is aimed at characterizing clustered DNA damage produced by DNA-damaging agents such as ionizing radiation and genotoxic chemicals. A fluorescent probe with an aminooxy group (AlexaFluor488) was used to label APs. We prepared a pUC19 plasmid with APs by heating under acidic conditions as a model for damaged DNA, and subsequently labeled the APs. We found that the observed fluorescence anisotropy ( r obs ) decreases as averaged AP density ( λ AP : number of APs per base pair) increases due to homo-FRET, and that the APs were randomly distributed. We applied this method to three DNA-damaging agents, 60 Co γ-rays, methyl methanesulfonate (MMS), and neocarzinostatin (NCS). We found that r obs – λ AP relationships differed significantly between MMS and NCS. At low AP density ( λ AP 60 Co γ-rays was similar to, but potentially more likely to occur than, random distribution. This simple method can be used to estimate mutagenicity of ionizing radiation and genotoxic chemicals.

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    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

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  2. [해외논문]   The role of human monoacylglycerol lipase (hMAGL) binding pocket in breakup of unsaturated phospholipid membranes   SCI SCIE

    Karageorgos, Ioannis (Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD, 20899, United States ) , Silin, Vitalii I. (Institute for Bioscience and Biotechnology Research, Rockville, MD, 20850, United States ) , Zvonok, Nikolai (Center for Drug Discovery, Northeastern University, Boston, MA, 02115, United States ) , Marino, John (Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD, 20899, United States ) , Janero, David R. (Center for Drug Discovery, Northeastern University, Boston, MA, 02115, United States ) , Makriyannis, Alexandros (Center for Drug Discovery, Northeastern University, Boston, MA, 02115, United States)
    Analytical biochemistry v.536 ,pp. 90 - 95 , 2017 , 0003-2697 ,

    초록

    Abstract Human monoacylglycerol lipase (hMAGL) plays a key role in homeostatic tuning of the endocannabinoid signaling system and supports aggressive tumorogenesis, making this enzyme a promising therapeutic target. hMAGL features a membrane-associated lid domain that regulates entry of endocannabinoid lipid substrates into the hydrophobic channel accessing the active site, likely from the membrane bilayer. The present work applied simultaneous surface plasmon resonance and electrochemical impedance spectroscopy measurements to show that, in absence of the substrate, hMAGL can remove phospholipid molecules from the membrane and, thereby, disintegrate pre-formed, intact, tethered phospholipid bilayer membrane mimetics (tBLMs) composed of unsaturated phosphatidylcholines. To probe the mechanism of hMAGL-induced on tBLMs compromise, we investigated the effect of wild type and mutant hMAGLs and hMAGL rendered catalytically inactive, as a function of concentration and in the presence of chemically distinct active-site inhibitors. Our data show that hMAGL's lid domain and hydrophobic substrate-binding pocket play important roles in hMAGL-induced bilayer lipid mobilization, whereas hydrolytic activity of the enzyme does not appear to be a factor.

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    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

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  3. [해외논문]   NADPH oxidase activity: Spectrophotometric determination of superoxide using pyrogallol red   SCI SCIE

    Corté (Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile ) , s-Rí (Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile ) , os, J. (Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile ) , Torres, M.J. (Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile ) , Campos-Bustamante, M.P. (Department of Pharmacological and Toxicological Chemistry, Faculty of Pharmaceutical and Chemical Sciences, Universidad de Chile, Chile ) , Romero-Parra, J. (Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile ) , Letelier, M.E. (Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile ) , Pessoa-Mahana, D. (Department of Pharmacy, Faculty of Chemistry, Pontificia Universidad Católica de Chile, Chile) , Chung, H. , Faú , ndez, M.
    Analytical biochemistry v.536 ,pp. 96 - 100 , 2017 , 0003-2697 ,

    초록

    Abstract A simple and fast spectrophotometric methodology able to quantify superoxide released by NADPH oxidase from differentiated promyelocytic leukaemia (HL-60) cells using pyrogallol red is described.The latter is based on the known stoichiometry of the reaction between superoxide and pyrogallol red and the inability of pyrogallol red to react with hydrogen peroxide. In addition, we developed a 96-wells microplate-based method able to determine NADPH oxidase activity. Using this method, we determined pharmacological properties of the NADPH oxidase inhibitors VAS2870 and diphenyleneiodonium and the obtained IC50 values were in good agreement with previous reported data. NOX2 is highly expressed in differentiated promyelocytic leukaemia cells, whereas other isoforms are not detected or expressed at low amounts. Likewise, this methodology may be a useful assay for NOX2 inhibitor screening. NADPH oxidases are involved in several physiological and pathological processes, rendering its pharmacological modulation an attractive research target. In this context, this simple assay can be used for NADPH oxidase inhibitor screening as well as aiding in the study of different biological conditions that involve NADPH oxidase activity. Highlights Neutrophil-like cells stimulated with PMA induce bleaching of PGR. PGR bleaching is induced by superoxide generated by NOX. PGR is a useful extracellular probe for the determination of NOX activity. Determinations were performed in a 96-wells microplate based method. Graphical abstract [DISPLAY OMISSION]

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    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

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    Fig. 1 이미지

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