MEASUREMENT OF SUPEROXIDE FORMATION BY MITOCHONDRIAL COMPLEX I OF YARROWIA LIPOLYTICA

S. Drose, Alexander Galkin, U. Brandt

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Complex I (NADH: ubiquinone oxidoreductase) is generally regarded as one of the major sources of mitochondrial reactive oxygen species (ROS). Mitochondrial membranes from the obligate aerobic yeast Yarrowia lipolytica, as well as the purified and reconstituted enzyme, can be used to measure complex I-dependent generation of superoxide (O-2(center dot-)). The use of isolated complex I excludes interference with other respiratory chain complexes and matrix enzymes during superoxide dismutase-sensitive reduction of acetylated cytochrome c. Alternately. hydrogen peroxide formation can be measured by the Amplex Red/horseradish peroxidase assay. Both methods allow the determination of complex I-generated ROS, depending on substrates (NADH, artificial ubiquinones), membrane potential, and active/deactive transition. ROS production by Yorrowia complex I in the
Original languageEnglish
Pages (from-to)475-490
Number of pages16
JournalMETHODS IN ENZYMOLOGY, VOL 456
Volume456
Issue numberA
DOIs
Publication statusPublished - 2009

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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