Complex I spectrophotometric assay in cultured cells: Detailed analysis of key factors

Complex I spectrophotometric assay in cultured cells: Detailed analysis of key factors

Autor Oliveira, Katia Klug Autor UNIFESP Google Scholar
Kiyomoto, Beatriz Hitomi Autor UNIFESP Google Scholar
Rodrigues, Andresa De Santi Autor UNIFESP Google Scholar
Tengan, Celia Harumi Autor UNIFESP Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Resumo 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.
Assunto Respiratory chain
Mitochondrial diseases
Complex I
Spectrophotometry
Mitochondria
Ubiquinone
NADH:ubiquinone reductase
Idioma Inglês
Financiador Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Data 2013-04-01
Publicado em Analytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 435, n. 1, p. 57-59, 2013.
ISSN 0003-2697 (Sherpa/Romeo, fator de impacto)
Editor Elsevier B.V.
Extensão 57-59
Fonte http://dx.doi.org/10.1016/j.ab.2012.12.002
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000315477600009
URI http://repositorio.unifesp.br/handle/11600/36152

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