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dc.contributor.authorRoberts, Gareth Aen
dc.contributor.authorCelik, Ayhanen
dc.contributor.authorHunter, Dominic JBen
dc.contributor.authorOst, Tobias WBen
dc.contributor.authorWhite, John Hen
dc.contributor.authorChapman, Stephen Ken
dc.contributor.authorTurner, Nicholas Jen
dc.contributor.authorFlitsch, Sabine Len
dc.coverage.spatial7en
dc.date.accessioned2005-01-31T17:00:38Z
dc.date.available2005-01-31T17:00:38Z
dc.date.issued2003-09-27
dc.identifier.citationJournal Of Biological Chemistry (2003), 278(49), 48914-48920
dc.identifier.uriDOI 10.1074/jbc.M309630200
dc.identifier.urihttp://www.jbc.org
dc.identifier.urihttp://hdl.handle.net/1842/663
dc.description.abstractP450 RhF from Rhodococcus sp. NCIMB 9784 is the first example of a new class of cytochrome P450 in which electrons are supplied by a novel, FMN- and Fe/S-containing, reductase partner in a fused arrangement. We have previously cloned the gene encoding the enzyme and shown it to comprise an N-terminal P450 domain fused to a reductase domain that displays similarity to the phthalate family of oxygenase reductase proteins. A reductase of this type had never previously been reported to interact with a cytochrome P450. In this report we describe the purification and partial characterization of P450 RhF. We show that the enzyme is selfsufficient in catalyzing the O-dealkylation of 7-ethoxycoumarin. The P450 RhF catalyzed O-dealkylation of 7-ethoxycoumarin is inhibited by several compounds that are known inhibitors of cytochrome P450. Presteady state kinetic analysis indicates that P450 RhF shows a 500-fold preference for NAPDH over NADH in terms of Kd value (6.6 -mu M versus 3.7 mM, respectively). Potentiometric studies show reduction potentials of -243 mV for the two-electron reduction of the FMN and -423 mV for the heme (in the absence of substrate).en
dc.format.extent223931 bytesen
dc.format.mimetypeapplication/pdfen
dc.language.isoen
dc.publisher© 2003 The American Society for Biochemistry and Molecular Biology, Inc.en
dc.subjectSelf-sufficient Cytochrome P450en
dc.subjectPrimary Structural Organizationen
dc.subjectFlavin-Domainen
dc.titleA Self-sufficient Cytochrome P450 with a Primary Structural Organization That Includes a Flavin Domain and a [2Fe-2S] Redox Centeren
dc.typeArticleen


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