title: Purification and characterization of cytochrome c oxidase from the insect trypanosomatid Crithidia fasciculata.
abstract: Cytochrome c oxidase was purified from the mitochondrial lysate of the insect trypanosomatid Crithidia fasciculata with the aid of a methyl hydrophobic interaction column in a rapid one-step procedure. The purified complex displayed all characteristics expected from a eukaryotic cytochrome c oxidase: the presence of CuA in electron paramagnetic resonance analysis, a characteristic 605 nm peak in reduced-minus-oxidized optical spectroscopy, and the capacity to efficiently oxidize homologous, but not heterologous, cytochrome c. Two-dimensional PAGE showed that C. fasciculata cytochrome c oxidase consists of at least 10 different subunits. N-terminal sequences were obtained from the six smallest subunits of the complex, one of them showing significant similarity to Neurospora crassa cytochrome c oxidase subunit V. The N-terminus of each of the four largest subunits was found to be blocked.
authors.affiliation: E.C. Slater Institute, University of Amsterdam, Academic Medical Centre, The Netherlands.
authors.forename: D
authors.initials: D
authors.lastname: Speijer
authors.forename: A O
authors.initials: AO
authors.lastname: Muijsers
authors.forename: H
authors.initials: H
authors.lastname: Dekker
authors.forename: A
authors.initials: A
authors.lastname: de Haan
authors.forename: C K
authors.initials: CK
authors.lastname: Breek
authors.forename: S P
authors.initials: SP
authors.lastname: Albracht
authors.forename: R
authors.initials: R
authors.lastname: Benne
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016831:Crithidia fasciculata / Q000201:enzymology*; D004578:Electron Spin Resonance Spectroscopy; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007313:Insecta / Q000469:parasitology; D007700:Kinetics; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry
chemical_list: D003576:Electron Transport Complex IV