title: EPR signals of cytochrome b558 purified from porcine neutrophils.
abstract: Cytochrome b558 of pig blood neutrophils was partially purified, and its EPR spectra were measured. The cytochrome b558 was solubilized from membranes with the detergent n-heptyl-beta-thioglucoside and purified by DEAE-Sepharose and heparin-Sepharose chromatographies. The small and large subunits of cytochrome b558 were detected on gel by immunoblotting. A solution of the purified, undenatured cytochrome b558 at 85-108 microM concentration was obtained. The concentrated cytochrome b558 showed an EPR signal at a g value of 3.26 with a bandwidth of 100 G at 10 K. Addition of 2 mM KCN had no effect on the low spin signal at g = 3.26 but caused disappearance of a minor high spin signal. The cyanide-insensitive signal at g = 3.26 disappeared completely on reduction with Na2S2O4. These results suggest that the g = 3.26 signal is characteristic of the low spin heme in cytochrome b558 of neutrophils.
authors.affiliation: Department of Inflammation Research, Tokyo Metropolitan Institute of Medical Science, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Miki
authors.forename: H
authors.initials: H
authors.lastname: Fujii
authors.forename: K
authors.initials: K
authors.lastname: Kakinuma
mesh_terms: D000818:Animals; D003573:Cytochrome b Group / Q000737:chemistry* / Q000302:isolation & purification; D004578:Electron Spin Resonance Spectroscopy; D008970:Molecular Weight; D019255:NADPH Oxidases; D009504:Neutrophils / Q000737:chemistry*; D013057:Spectrum Analysis; D013552:Swine
chemical_list: D003573:Cytochrome b Group; C036888:cytochrome b558; D019255:NADPH Oxidases