title: Isolation and characterization of the flavin-binding domain of flavocytochrome b2 expressed independently in Escherichia coli.
abstract: Flavocytochrome b2 consists of two distinct domains. The N-terminal domain contains protohaem IX and the larger, C-terminal domain contains flavin mononucleotide (FMN). We describe here the isolation of the flavin-binding domain expressed in Escherichia coli independent of the cytochrome domain. The isolated domain is an efficient lactate dehydrogenase with ferricyanide as electron acceptor but reduces cytochrome c, the physiological oxidant for flavocytochrome b2, extremely poorly; electron transfer from the flavin-binding domain to the separately expressed cytochrome domain is undetectable. FMN reduction by lactate occurs as a single exponential process in the isolated flavin-binding domain, in contrast to the biphasic kinetics observed with native flavocytochrome b2.
authors.affiliation: Department of Chemistry, University of Edinburgh, Scotland, UK.
authors.forename: A
authors.initials: A
authors.lastname: Balme
authors.forename: C E
authors.initials: CE
authors.lastname: Brunt
authors.forename: R L
authors.initials: RL
authors.lastname: Pallister
authors.forename: S K
authors.initials: SK
authors.lastname: Chapman
authors.forename: G A
authors.initials: GA
authors.lastname: Reid
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005415:Flavins / Q000378:metabolism*; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D042844:L-Lactate Dehydrogenase (Cytochrome); D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010084:Oxidation-Reduction; D011485:Protein Binding
chemical_list: D005415:Flavins; D009838:Oligodeoxyribonucleotides; D007770:L-Lactate Dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome)