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Title: Complex I spectrophotometric assay in cultured cells: Detailed analysis of key factors
Authors: Oliveira, Katia Klug [UNIFESP]
Kiyomoto, Beatriz Hitomi [UNIFESP]
Rodrigues, Andresa De Santi [UNIFESP]
Tengan, Celia Harumi [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
Keywords: Respiratory chain
Mitochondrial diseases
Complex I
NADH:ubiquinone reductase
Issue Date: 1-Apr-2013
Publisher: Elsevier B.V.
Citation: Analytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 435, n. 1, p. 57-59, 2013.
Abstract: 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.
ISSN: 0003-2697
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Appears in Collections:Em verificação - Geral

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