title: Amperometric assay for aldolase activity: antibody-catalyzed ferrocenylamine formation.
abstract: Screening of new catalysts for aldolase activity is a major task in bioorganic chemistry. For this purpose, fast and convenient methods are required for the detection of the catalysts. We have developed the first amperometric assay for aldol or retro-aldol catalytic activity. A new ferrocene-aldol derivative was synthesized with redox activity significantly different from that of ferrocenylamine. It was shown that the reaction between aldolase antibody 38C2 and a ferrocene-aldol substrate generated free ferrocenylamine, which could be detected and quantified by simple electrochemical measurement. The amperometric assay was applied to perform a Michaelis-Menten analysis of catalytic antibody 38C2 in order to determine the enzymatic kinetic parameters.
authors.affiliation: School of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel-Aviv University, Tel Aviv 69978 Israel.
authors.forename: Amit
authors.initials: A
authors.lastname: Sagi
authors.forename: Judith
authors.initials: J
authors.lastname: Rishpon
authors.forename: Doron
authors.initials: D
authors.lastname: Shabat
mesh_terms: D000906:Antibodies; D002384:Catalysis; D004563:Electrochemistry / Q000379:methods*; D004566:Electrodes; D005296:Ferrous Compounds / Q000032:analysis / Q000138:chemical synthesis*; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis* / Q000378:metabolism
chemical_list: D000906:Antibodies; D005296:Ferrous Compounds; D005634:Fructose-Bisphosphate Aldolase