title: Optical, EPR and Mössbauer spectroscopic studies on the NO derivatives of cytochrome cd1 from Thiobacillus denitrificans.
abstract: We have used optical, EPR and Mössbauer spectroscopies to study the formation of heme-NO complex upon the addition of nitrite to reduced cytochrome cd1 from Thiobacillus denitrificans. The reduced d1 heme binds NO under both alkaline and acidic conditions, but the binding of NO to the reduced c heme was strongly pH-dependent. The Mössbauer data showed unambiguously that at pH 7.6 the c heme does not complex NO, whereas at pH 5.8 approximately half of the reduced c heme binds NO. This observation was confirmed by EPR studies, which showed that the spin concentration of the heme-NO EPR signal increased from 2 spins/molecule at pH 8.0 to approximately 3 spins/molecule at pH 5.8. Optical absorption study also showed strong pH dependence in the binding of NO to the reduced c heme. We have also analyzed the Mössbauer spectra of the ferrous d1 heme-NO complex using a spin-Hamiltonian formalism. The magnetic hyperfine coupling tensor was found to be consistent with the unpaired electron residing on a sigma orbital.
authors.affiliation: Department of Microbiology, University of Georgia, Athens.
authors.forename: M C
authors.initials: MC
authors.lastname: Liu
authors.forename: B H
authors.initials: BH
authors.lastname: Huynh
authors.forename: W J
authors.initials: WJ
authors.lastname: Payne
authors.forename: H D
authors.initials: HD
authors.lastname: Peck
authors.forename: D V
authors.initials: DV
authors.lastname: Dervartanian
authors.forename: J
authors.initials: J
authors.lastname: Legall
mesh_terms: D003574:Cytochrome c Group; D003580:Cytochromes / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D009572:Nitrite Reductases; D010084:Oxidation-Reduction; D011485:Protein Binding; D013053:Spectrophotometry; D013057:Spectrum Analysis; D013855:Thiobacillus / Q000378:metabolism*
chemical_list: D003574:Cytochrome c Group; D003580:Cytochromes; D009569:Nitric Oxide; D006418:Heme; C019315:cytochrome cd1; D009572:Nitrite Reductases