title: Spectroscopic studies on an oxygen-binding haemoglobin-like flavohaemoprotein from Escherichia coli.
abstract: The Escherichia coli haemoglobin-like flavohaemoprotein (Hmp) has been purified to near homogeneity using two chromatographic steps. The prosthetic groups are identified as FAD and protohaem IX. SDS/PAGE has indicated a molecular mass of 44 kDa for the monomeric protein consistent with the amino-acid sequence deduced from the hmp+ gene. The protein, as isolated, is in the Fe(III) state, exhibiting absorbance maxima at 403.5, 540 (shoulder) and 627 nm. The ferrous and carbonmonoxyferrous states resemble those of haemoglobin, showing maxima at 431.5 and 558 nm, and 421, 542 and 566 nm respectively. Upon aerobic addition of NAD(P)H, the ferric state is reduced to the oxygenated Fe(II) state, characterized by maxima at 413, 544 and 580 nm. This oxy form is not stable and slowly decays to the ferric state. Addition of dithionite and nitrite to the ferric protein results in the formation of a nitrosyl complex, whose e.p.r. characteristics indicate that the b-type haem is attached to the protein through a nitrogenous ligand, probably originating from a histidine residue.
authors.affiliation: Division of Biosphere Sciences, King's College, University of London, U.K.
authors.forename: N
authors.initials: N
authors.lastname: Ioannidis
authors.forename: C E
authors.initials: CE
authors.lastname: Cooper
authors.forename: R K
authors.initials: RK
authors.lastname: Poole
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D004093:Dihydropteridine Reductase / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli / Q000737:chemistry; D005290:Ferric Compounds / Q000737:chemistry; D005420:Flavoproteins / Q000737:chemistry*; D006418:Heme / Q000737:chemistry; D006420:Hemeproteins / Q000737:chemistry*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry; D013057:Spectrum Analysis
chemical_list: D001426:Bacterial Proteins; D005290:Ferric Compounds; D005420:Flavoproteins; D006420:Hemeproteins; D006418:Heme; D004093:Dihydropteridine Reductase; D010100:Oxygen