title: Quinone- and nitroreductase reactions of Thermotoga maritima peroxiredoxin-nitroreductase hybrid enzyme.
abstract: Thermotoga maritima peroxiredoxin-nitroreductase hybrid enzyme (Prx-NR) consists of a FMN-containing nitroreductase (NR) domain fused to a peroxiredoxin (Prx) domain. These domains seem to function independently as no electron transfer occurs between them. The reduction of quinones and nitroaromatics by NR proceeded in a two-electron manner, and follows a 'ping-pong' scheme with sometimes pronounced inhibition by quinone substrate. The comparison of steady- and presteady-state kinetic data shows that in most cases, the oxidative half-reaction may be rate-limiting in the catalytic cycle of NR. The enzyme was inhibited by dicumarol, a classical inhibitor of oxygen-insensitive nitroreductases. The reduction of quinones and nitroaromatic compounds by Prx-NR was characterized by the linear dependence of their reactivity (logk(cat)/K(m)) on their single-electron reduction potentials E(7)(1), while the reactivity of quinones markedly exceeded the one with nitroaromatics. It shows that NR lacks the specificity for the particular structure of these oxidants, except their single-electron accepting potency and the rate of electron self-exchange. It points to the possibility of a single-electron transfer step in a net two-electron reduction of quinones and nitroaromatics by T. maritima Prx-NR, and to a significant diversity of the structures of flavoenzymes which may perform the two-electron reduction of quinones and nitroaromatics.
authors.affiliation: Institute of Biochemistry of Vilnius University, Mokslininkų 12, LT-08662 Vilnius, Lithuania.
authors.forename: Žilvinas
authors.initials: Ž
authors.lastname: Anusevičius
authors.forename: Lina
authors.initials: L
authors.lastname: Misevičienė
authors.forename: Jonas
authors.initials: J
authors.lastname: Šarlauskas
authors.forename: Nicolas
authors.initials: N
authors.lastname: Rouhier
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Jacquot
authors.forename: Narimantas
authors.initials: N
authors.lastname: Čėnas
mesh_terms: D005486:Flavin Mononucleotide / Q000378:metabolism; D007700:Kinetics; D009574:Nitro Compounds / Q000378:metabolism; D009601:Nitroreductases / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D054464:Peroxiredoxins / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011809:Quinones / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D020124:Thermotoga maritima / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism
chemical_list: D009574:Nitro Compounds; D011809:Quinones; D011993:Recombinant Fusion Proteins; D005486:Flavin Mononucleotide; D054464:Peroxiredoxins; D009601:Nitroreductases