title: Kinetic characterization of antibody-catalyzed insertion of a metal ion into porphyrin.
abstract: The insertion of a copper (II) ion into mesoporphyrin by a monoclonal catalytic antibody has been investigated kinetically by measuring the increase in Soret absorbance due to the production of copper (II)-mesoporphyrin. The initial rate of the reaction showed saturation kinetics as a function of the mesoporphyrin concentration, while it increased linearly with an increase in the copper (II) concentration. Based on observations, a scheme for the reaction was proposed: mesoporphyrin binds to the antibody to form a complex, and copper (II) binds to the complex to yield copper (II)-mesoporphyrin. Kinetic parameters for the respective steps were estimated, and the thermodynamic parameters were calculated. The binding of mesoporphyrin to the antibody was endothermic and entropically driven. This implies that hydrophobic interactions are an important factor in the binding. Free energy profiles for the antibody-catalyzed and uncatalyzed reactions were drawn by use of the obtained thermodynamic parameter values. The results demonstrate that the rate acceleration by, the antibody is ascribable to transition-state stabilization, and suggest that the structure of mesoporphyrin in the complex is more distorted than that of free mesoporphyrin.
authors.affiliation: Department of Biosciences, School of Science, Kitasato University, Kanagawa.
authors.forename: Y
authors.initials: Y
authors.lastname: Kawamura-Konishi
authors.forename: N
authors.initials: N
authors.lastname: Hosomi
authors.forename: S
authors.initials: S
authors.lastname: Neya
authors.forename: S
authors.initials: S
authors.lastname: Sugano
authors.forename: N
authors.initials: N
authors.lastname: Funasaki
authors.forename: H
authors.initials: H
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D017151:Antibodies, Catalytic / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D002384:Catalysis; D003300:Copper / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008652:Mesoporphyrins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D013816:Thermodynamics
chemical_list: D017151:Antibodies, Catalytic; D000911:Antibodies, Monoclonal; D008652:Mesoporphyrins; C031468:mesoporphyrin IX; D003300:Copper