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

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dc.contributor.authorOliveira, Katia Klug [UNIFESP]
dc.contributor.authorKiyomoto, Beatriz Hitomi [UNIFESP]
dc.contributor.authorRodrigues, Andresa De Santi [UNIFESP]
dc.contributor.authorTengan, Celia Harumi [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T14:31:30Z
dc.date.available2016-01-24T14:31:30Z
dc.date.issued2013-04-01
dc.description.abstractThe 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.en
dc.description.affiliationUniversidade Federal de São Paulo, Escola Paulista Med, Dept Neurol & Neurosurg, BR-04039032 São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Escola Paulista Med, Dept Neurol & Neurosurg, BR-04039032 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.format.extent57-59
dc.identifierhttp://dx.doi.org/10.1016/j.ab.2012.12.002
dc.identifier.citationAnalytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 435, n. 1, p. 57-59, 2013.
dc.identifier.doi10.1016/j.ab.2012.12.002
dc.identifier.issn0003-2697
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/36152
dc.identifier.wosWOS:000315477600009
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofAnalytical Biochemistry
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectRespiratory chainen
dc.subjectMitochondrial diseasesen
dc.subjectComplex Ien
dc.subjectSpectrophotometryen
dc.subjectMitochondriaen
dc.subjectUbiquinoneen
dc.subjectNADH:ubiquinone reductaseen
dc.titleComplex I spectrophotometric assay in cultured cells: Detailed analysis of key factorsen
dc.typeinfo:eu-repo/semantics/article
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