title: Models of the low-spin iron(III) hydroperoxide intermediate of heme oxygenase: magnetic resonance evidence for thermodynamic stabilization of the d(xy) electronic state at ambient temperatures.
abstract: The (13)C pulsed ENDOR and NMR study of [meso-(13)C-TPPFe(OCH(3))(OO(t)Bu)](-) performed in this work shows that although the unpaired electron in low-spin ferrihemes containing a ROO(-) ligand resides in a d(pi) orbital at 8 K, the d(xy) electron configuration is favored at physiological temperatures. The variable temperature NMR spectra indicate a dynamic situation in which a heme with a d(pi) electron configuration and planar porphyrinate ring is in equilibrium with a d(xy) electron configuration that has a ruffled porphyrin ring. Because of the similarity in the EPR spectra of the hydroperoxide complexes of heme oxygenase, cytochrome P450, and the model heme complex reported herein, it is possible that these two electron configurations and ring conformations may also exist in equilibrium in the enzymatic systems. The ruffled porphyrinate ring would aid the attack of the terminal oxygen of the hydroperoxide intermediate of heme oxygenase (HO) on the meso-carbon, and the large spin density at the meso-carbons of a d(xy) electron configuration heme suggests the possibility of a radical mechanism for HO. The dynamic equilibrium between the ruffled (d(xy)) and planar (d(pi)) conformers observed in the model complexes also suggests that a flexible heme binding cavity may be an important structural motif for heme oxygenase activity.
authors.affiliation: Department of Chemistry, Oklahoma State University, Stillwater, Oklahoma 74078-3071, USA. Rivera@okstate.edu
authors.forename: Mario
authors.initials: M
authors.lastname: Rivera
authors.forename: Gregori A
authors.initials: GA
authors.lastname: Caignan
authors.forename: Andrei V
authors.initials: AV
authors.lastname: Astashkin
authors.forename: Arnold M
authors.initials: AM
authors.lastname: Raitsimring
authors.forename: Tatjana Kh
authors.initials: TKh
authors.lastname: Shokhireva
authors.forename: F Ann
authors.initials: FA
authors.lastname: Walker
mesh_terms: D002247:Carbon Isotopes; D004578:Electron Spin Resonance Spectroscopy; D004583:Electrons; D005290:Ferric Compounds / Q000737:chemistry*; D006419:Heme Oxygenase (Decyclizing) / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008665:Metalloporphyrins / Q000737:chemistry; D008956:Models, Chemical; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D011487:Protein Conformation; D013696:Temperature; D013816:Thermodynamics; D020122:tert-Butylhydroperoxide / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D005290:Ferric Compounds; D008665:Metalloporphyrins; D020122:tert-Butylhydroperoxide; D006419:Heme Oxygenase (Decyclizing)