title: Evaluation of the cytochrome b(5)/cytochrome b(5) reductase pathway.
abstract: NADH cytochrome b(5) reductase (b(5)R; EC 1.6.2.2; Diaphorase I; NADH: ferricytochrome b(5) oxidoreductase) is an FAD-containing protein, which, along with the hemoprotein cytochrome b(5) (cyt b(5)), mediates electron transfer from NADH to fatty acid desaturases, P450 oxidases, methemoglobin, and ascorbyl free radical. In addition, b(5)R and cyt b(5) can directly catalyze the reduction of hydroxylamine and amidoxime metabolites. This unit provides protocols for measuring the activity and mRNA expression of the cytochrome b(5)/cytochrome b(5) reductase pathway, and for obtaining heterologous expression and purification of the soluble forms of each protein.
authors.affiliation: University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Lauren A
authors.initials: LA
authors.lastname: Trepanier
authors.forename: Sunil U
authors.initials: SU
authors.lastname: Bajad
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Kurian
mesh_terms: D001483:Base Sequence; D002478:Cells, Cultured; D015786:Cytochromes b5 / Q000378:metabolism*; D017931:DNA Primers; D006801:Humans; D008706:Methemoglobin / Q000378:metabolism; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D008706:Methemoglobin; D015786:Cytochromes b5