Navegando por Palavras-chave "Complex I"

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    Complex I spectrophotometric assay in cultured cells: Detailed analysis of key factors
    (Elsevier B.V., 2013-04-01) Oliveira, Katia Klug [UNIFESP]; Kiyomoto, Beatriz Hitomi [UNIFESP]; Rodrigues, Andresa De Santi [UNIFESP]; Tengan, Celia Harumi [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    The diagnosis of mitochondrial encephalomyopathies caused by complex I (C-I) deficiency relies mainly on the spectrophotometric C-I assay. Considered difficult, this assay lacks reliability and has high nonspecific activity. We studied the key factors of this assay in cultured cells (cybrid and fibroblast): ubiquinone analogues, rotenone inhibition to determine specific activity, and mode of permeabilization of mitochondria] membranes. We showed that ubiquinone 1 allows a more stable and reliable assay, better results were obtained with rotenone inhibition done in the same assay, and mitochondria] permeability was improved just by freeze/thawing the sample prior to the assay. (C) 2012 Elsevier Inc. All rights reserved.