title: P450 oxidoreductase: genotyping, expression, purification of recombinant protein, and activity assessments of wild-type and mutant protein.
abstract: P450 oxidoreductase (POR) is the flavoprotein that transfers electrons from NADPH to microsomal cytochrome P450 enzymes and to some other proteins. Protocols for genotyping human POR for common polymorphisms are described. Expression in E. coli of recombinant human POR, its purification, and different methods of assessing the effect of amino-acid sequence variants of POR on the activity of various cytochromes P450 are also described.
authors.affiliation: Department of Pediatrics, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Vishal
authors.initials: V
authors.lastname: Agrawal
authors.forename: Walter L
authors.initials: WL
authors.lastname: Miller
mesh_terms: D002462:Cell Membrane / Q000235:genetics; D004247:DNA / Q000097:blood / Q000235:genetics; D057075:Enzyme Assays / Q000379:methods*; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics; D015870:Gene Expression; D060005:Genotyping Techniques; D006801:Humans; D008861:Microsomes / Q000201:enzymology; D009154:Mutation; D009251:NADPH-Ferrihemoprotein Reductase / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D009532:Nickel / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D009532:Nickel; D004247:DNA; D009251:NADPH-Ferrihemoprotein Reductase