title: Inhibition of lignin peroxidase H2 by sodium azide.
abstract: The oxidation of veratryl alcohol (3,4-dimethoxybenzyl alcohol) by lignin peroxidase H2 from Phanerochaete chrysosporium and H2O2 was strongly inhibited by sodium azide. Inhibition was competitive with respect to veratryl alcohol (Ki = 1-2 microM) and uncompetitive with respect to H2O2. In contrast, sodium azide bound to the native enzyme at pH 6.0 with an apparent dissociation constant (KD) of 126 mM. Formation of azidyl radicals was detected by ESR spin trapping techniques. The enzymes is nearly completely inactivated in four turnovers. The H2O2-activated enzyme intermediate (compound I) reacted with sodium azide to form a new species rather than be reduced to the enzyme intermediate compound II. The new species has absorption maxima at 418, 540, and 570 nm, suggesting the formation of a ferrous-lignin peroxidase-NO complex. Confirmation of this assignment was obtained by low-temperature ESR spectroscopy. An identical complex could be simulated by the addition of nitrite to the reduced enzyme. The enzyme intermediate compound II is readily reduced by sodium azide to native enzyme with essentially no loss of activity.
authors.affiliation: Biotechnology Center, Utah State University, Logan 84322-4700.
authors.forename: H
authors.initials: H
authors.lastname: Tuisel
authors.forename: T A
authors.initials: TA
authors.lastname: Grover
authors.forename: J R
authors.initials: JR
authors.lastname: Lancaster
authors.forename: J A
authors.initials: JA
authors.lastname: Bumpus
authors.forename: S D
authors.initials: SD
authors.lastname: Aust
mesh_terms: D000363:Agaricales / Q000201:enzymology*; D001386:Azides / Q000494:pharmacology*; D001592:Benzyl Alcohols / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D010544:Peroxidases / Q000037:antagonists & inhibitors*; D019810:Sodium Azide; D013053:Spectrophotometry
chemical_list: D001386:Azides; D001592:Benzyl Alcohols; D019810:Sodium Azide; D010544:Peroxidases; C042858:lignin peroxidase; C042197:veratryl alcohol