title: Design of a novel P450: a functional bacterial-human cytochrome P450 chimera.
abstract: We report the construction of a functional chimera from approximately 50% bacterial (cytosolic) cytochrome P450cam and 50% mammalian (membrane-bound) cytochrome P450 2C9. The chimeric protein shows a reduced CO-difference spectrum absorption at 446 nm, and circular dichroism spectra indicate that the protein is globular. The protein is soluble and catalyzes the oxidation of 4-chlorotoluene using molecular oxygen and reducing equivalents from bacterial putidaredoxin and putidaredoxin reductase. This chimera provides a novel method for addressing structure-function issues and may prove useful in the design of oxidants for benign and stereospecific synthesis, as well as catalysts for bioremediation of polluted areas. Furthermore, these results provide the first evidence that bacterial P450 enzymes and mammalian P450 enzymes are likely to share a common tertiary structure.
authors.affiliation: Department of Biochemistry, University of Rochester, New York 14642, USA.
authors.forename: M
authors.initials: M
authors.lastname: Shimoji
authors.forename: H
authors.initials: H
authors.lastname: Yin
authors.forename: L
authors.initials: L
authors.lastname: Higgins
authors.forename: J P
authors.initials: JP
authors.lastname: Jones
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D001665:Binding Sites; D002384:Catalysis; D002942:Circular Dichroism; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000138:chemical synthesis* / Q000235:genetics*; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D006801:Humans; D008958:Models, Molecular; D010100:Oxygen / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000138:chemical synthesis* / Q000378:metabolism*; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D003577:Cytochrome P-450 Enzyme System; D010100:Oxygen