title: Effects of Active-Site Modification and Quaternary Structure on the Regioselectivity of Catechol-O-Methyltransferase.
abstract: Catechol-O-methyltransferase (COMT), an important therapeutic target in the treatment of Parkinson's disease, is also being developed for biocatalytic processes, including vanillin production, although lack of regioselectivity has precluded its more widespread application. By using structural and mechanistic information, regiocomplementary COMT variants were engineered that deliver either meta- or para-methylated catechols. X-ray crystallography further revealed how the active-site residues and quaternary structure govern regioselectivity. Finally, analogues of AdoMet are accepted by the regiocomplementary COMT mutants and can be used to prepare alkylated catechols, including ethyl vanillin.
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Brian J C
authors.initials: BJ
authors.lastname: Law
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Bennett
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Mark L
authors.initials: ML
authors.lastname: Thompson
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Colin
authors.initials: C
authors.lastname: Levy
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Shepherd
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: David
authors.initials: D
authors.lastname: Leys
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK. Jason.micklefield@manchester.ac.uk.
authors.forename: Jason
authors.initials: J
authors.lastname: Micklefield
mesh_terms: D020134:Catalytic Domain; D002394:Catechol O-Methyltransferase / Q000737:chemistry / Q000378:metabolism*; D018360:Crystallography, X-Ray; D020836:Protein Structure, Quaternary
chemical_list: D002394:Catechol O-Methyltransferase
keywords: biocatalysis; enzyme structure; methyltransferases; protein engineering; regioselectivity