nad a22 4500 551414162 20181214071544.0 cr unu---uuuuu 181214e20000101xx s 000 0 eng 10.5167/uzh-156296 doi 10.1385/abab:83:1-3:183 doi (ZORA)oai:www.zora.uzh.ch:156296 Gramatikova Svetlana I. Pyridoxal-5'-phosphate-dependent catalytic antibodies [Elektronische Daten] [Svetlana I Gramatikova, Philipp Christen] openAccess eu-repo Cofactors—i.e., metal ions and coenzymes—extend the catalytic scope of enzymes and might have been among the first biological catalysts. They may be expected to efficiently extend the catalytic potential of antibodies. Monoclonal antibodies (MAbs) against Nα-phosphopyridoxyl-l-lysine were screened for 1) binding of 5′-phosphopyridoxyl amino acids, 2) binding of the planar Schiff base of pyridoxal-5′-phosphate (PLP) and amino acids, the first intermediate of all PLP-dependent reactions, and 3), catalysis of the PLP-dependent α, β-elimination reaction with β-chloro-D/L-alanine. Antibody 15A9 fulfilled all criteria and was also found to catalyze the cofactor-dependent transamination reaction of hydrophobic D-amino acids and oxo acids (k′ cat=0.42 min−1 with D-alanine at 25°C). No other reactions with either D- or L-amino acids were detected. PLP markedly contributes to catalytic effecacy—it is a 104 times more efficient acceptor of the amino group than pyruvate. The antibody ensures reaction specificity, stereospecificity, and substrate specificity, and further accelerates the transamination reaction (k′ cat(Ab)/k′ cat(PLP)=5×103). The successive screening steps simulate the selection criteria that might have been operative in the evolution of protein-assisted psyridoxal catalysis 142-005 zora Christen Philipp joint author Applied Biochemistry and Biotechnology 83(1-3):183-194 https://www.zora.uzh.ch/id/eprint/156296/1/ZORA_NL_156296.pdf text/html WWW-Backlink auf das Repository 1 Journal Article PeerReviewed zora ZORA 856 40 https://www.zora.uzh.ch/id/eprint/156296/1/ZORA_NL_156296.pdf text/html WWW-Backlink auf das Repository ZORA 100 1- Gramatikova Svetlana I. ZORA 700 1- Christen Philipp joint author ZORA 773 0- Applied Biochemistry and Biotechnology 83(1-3):183-194 SWISSBIB 295298537 BK010053 XK010053 XK010000 ZORA ZORA ZORA Journal Article openAccess