title: The conformation of P450cam in complex with putidaredoxin is dependent on oxidation state.
abstract: Double electron-electron resonance (DEER) spectroscopy was used to determine the conformational state in solution for the heme monooxygenase P450cam when bound to its natural redox partner, putidaredoxin (Pdx). When oxidized Pdx was titrated into substrate-bound ferric P450cam, the enzyme shifted from the closed to the open conformation. In sharp contrast, however, the enzyme remained in the closed conformation when ferrous-CO P450cam was titrated with reduced Pdx. This result fully supports the proposal that binding of oxidized Pdx to P450cam opposes the open-to-closed transition induced by substrate binding. However, the data strongly suggest that in solution, binding of reduced Pdx to P450cam does not favor the open conformation. This supports a model in which substrate recognition is associated with the open-to-closed transition and electron transfer from Pdx occurs in the closed conformation. The opening of the enzyme in the ferric-hydroperoxo state following electron transfer from Pdx would provide for efficient O2 bond activation, substrate oxidation, and product release.
authors.affiliation: Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, USA.
authors.forename: William K
authors.initials: WK
authors.lastname: Myers
authors.forename: Young-Tae
authors.initials: YT
authors.lastname: Lee
authors.forename: R David
authors.initials: RD
authors.lastname: Britt
authors.forename: David B
authors.initials: DB
authors.lastname: Goodin
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D019475:Camphor 5-Monooxygenase / Q000737:chemistry / Q000378:metabolism*; D005288:Ferredoxins / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D011485:Protein Binding; D011487:Protein Conformation; D016958:Pseudomonas putida / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D005288:Ferredoxins; C100137:putidaredoxin; D019475:Camphor 5-Monooxygenase
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