title: Enzymatic kinetics of the quinol peroxidase of an aggressive periodontopathic bacterium.
abstract: Aggregatibacter actinomycetemcomitans is an oral pathogen for aggressive periodontitis, and encodes a triheme c-containing membrane-bound enzyme, quinol peroxidase (QPO) that catalyzes peroxidase activity using quinol in the respiratory chain. In the previous work, we have characterized recombinant QPO purified from the membrane fraction of Escherichia coli harboring a plasmid containing QPO gene. Irreversible inactivation of QPO by high concentration of H2O2 exhibited pseudo-first order kinetics. Analysis of initial-rate kinetics of QPO may suggest that enzyme catalytic mechanism is explained by a Ping Pong Bi Bi system rather than sequential systems. In addition, the redox reactions of cytochrome c in the presence of several values of [Q1H2]/[Q1] were at equilibrium, and only about 2/3 of the cytochrome c of QPO is reduced at high ratios of [Q1H2]/[Q1]. These results indicated that one of the three heme c moieties of QPO is maintained in an oxidized form even at increased ratios of [Q1H2]/[Q1], suggesting that QPO is reduced in the absence of H2O2 and only two of the three heme c moieties are reduced in the presence of high concentration of the Q1H2. Product inhibition of QPO accorded with our theoretical model for the reaction mechanism. Considered together, the enzymatic kinetics data for QPO confirm the Ping Pong Bi Bi system.
authors.affiliation: Department of Microbiology, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.
authors.forename: Tasuku
authors.initials: T
authors.lastname: Abe
authors.affiliation: Department of Microbiology, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.
authors.forename: Taketo
authors.initials: T
authors.lastname: Kawarai
authors.affiliation: Department of Microbiology, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Takahashi
authors.affiliation: Department of Microbiology, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Konishi
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans / Q000201:enzymology*; D006873:Hydroquinones / Q000378:metabolism*; D007700:Kinetics; D010544:Peroxidases / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D006873:Hydroquinones; D011994:Recombinant Proteins; D010544:Peroxidases
keywords: Ping Pong Bi Bi; enzymatic kinetics; hydrogen peroxide; peroxidase; respiratory chain