title: Escherichia coli flavohemoglobin is an efficient alkylhydroperoxide reductase.
abstract: Escherichia coli flavohemoglobin (HMP) is shown to be capable of catalyzing the reduction of several alkylhydroperoxide substrates into their corresponding alcohols using NADH as an electron donor. In particular, HMP possesses a high catalytic activity and a low Km toward cumyl, linoleic acid, and tert-butyl hydroperoxides, whereas it is a less efficient hydrogen peroxide scavenger. An analysis of UV-visible spectra during the stationary state reveals that at variance with classical peroxidases, HMP turns over in the ferrous state. In particular, an iron oxygen adduct intermediate whose spectrum is similar to that reported for the oxo-ferryl derivative in peroxidases (Compound II), has been identified during the catalysis of hydrogen peroxide reduction. This finding suggests that hydroperoxide cleavage occurs upon direct binding of a peroxide oxygen atom to the ferrous heme iron. Competitive inhibition of the alkylhydroperoxide reductase activity by carbon monoxide has also been observed, thus confirming that heme iron is directly involved in the catalytic mechanism of hydroperoxide reduction. The alkylhydroperoxide reductase activity taken together with the unique lipid binding properties of HMP suggests that this protein is most likely involved in the repair of the lipid membrane oxidative damage generated during oxidative/nitrosative stress.
authors.affiliation: CNR Istituto di Biologia e Patologia Molecolare, Department of Biochemical Sciences, University La Sapienza, 00185 Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Bonamore
authors.forename: Patrizia
authors.initials: P
authors.lastname: Gentili
authors.forename: Andrea
authors.initials: A
authors.lastname: Ilari
authors.forename: M Eugenia
authors.initials: ME
authors.lastname: Schininà
authors.forename: Alberto
authors.initials: A
authors.lastname: Boffi
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002384:Catalysis; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000378:metabolism; D006420:Hemeproteins / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D010544:Peroxidases / Q000737:chemistry / Q000378:metabolism*; D054464:Peroxiredoxins; D010741:Phospholipases A / Q000378:metabolism; D011487:Protein Conformation; D013053:Spectrophotometry
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006420:Hemeproteins; C094007:hmpX protein, Bacteria; D010544:Peroxidases; D054464:Peroxiredoxins; D010741:Phospholipases A