title: Pyridoxal 5'-phosphate-dependent catalytic antibody.
abstract: Cofactors might efficiently extend the catalytic potential of antibodies. Monoclonal antibodies against Nalpha-phosphopyridoxyl-L-lysine were screened for: 1) binding of 5'-phosphopyridoxyl amino acids, 2) binding of Schiff base of pyridoxal 5'-phosphate (PLP) and amino acids, the first intermediate of all PLP-dependent reactions, and 3) catalysis of PLP-dependent alpha,beta-elimination with beta-chloro-D/L-alanine. All three criteria were met by antibody 15A9. Further analysis for PLP-dependent reactions showed that this antibody catalyzes the cofactor-dependent transamination of hydrophobic D-amino acids and oxo acids (kcat' = 0.42 min-1 with D-alanine). No other reactions with either D- or L-amino acids were taking place. PLP markedly contributes to catalytic efficacy, being a 10(4) times more efficient acceptor of the amino group than pyruvate. The antibody further accelerates the reaction (kcat(antibody)'/kcat(PLP)' = 5 x 10(3) with D-alanine as substrate) and ensures reaction specificity, stereospecificity, as well as limited substrate specificity.
authors.affiliation: Biochemisches Institut der Universität Zürich, CH-8057 Zürich, Switzerland.
authors.forename: S I
authors.initials: SI
authors.lastname: Gramatikova
authors.forename: P
authors.initials: P
authors.lastname: Christen
mesh_terms: D000818:Animals; D017151:Antibodies, Catalytic / Q000737:chemistry*; D000941:Antigens; D001667:Binding, Competitive; D006241:Haptens; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011732:Pyridoxal Phosphate / Q000737:chemistry* / Q000276:immunology; D012545:Schiff Bases
chemical_list: D017151:Antibodies, Catalytic; D000941:Antigens; D006241:Haptens; D012545:Schiff Bases; D011732:Pyridoxal Phosphate