1. Enzyme Activation with a Synthetic Catalytic Co-enzyme Intermediate: Nucleotide Methylation by Flavoenzymes
- Author
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Djemel Hamdane, Vincent Guérineau, Charles Bou-Nader, David Cornu, Thibault Fogeron, Marc Fontecave, Chaire Chimie des processus biologiques, Laboratoire de Chimie des Processus Biologiques (LCPB), Collège de France (CdF (institution))-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Collège de France (CdF (institution))-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut Européen des membranes (IEM), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM), Institut de Chimie des Substances Naturelles (ICSN), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), and Université Montpellier 2 - Sciences et Techniques (UM2)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0301 basic medicine ,Models, Molecular ,reaction intermediates ,Stereochemistry ,[SDV]Life Sciences [q-bio] ,010402 general chemistry ,01 natural sciences ,Methylation ,Catalysis ,Cofactor ,Nucleic acid metabolism ,chemistry.chemical_compound ,Enzyme activator ,03 medical and health sciences ,RNA, Transfer ,Flavins ,Thermotoga maritima ,Uracil ,SICAPS ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,tRNA Methyltransferases ,biology ,Chemistry ,Nucleotides ,RNA ,General Chemistry ,General Medicine ,Thymidylate Synthase ,0104 chemical sciences ,Enzyme Activation ,030104 developmental biology ,Enzyme ,Catalytic cycle ,Biochemistry ,Transfer RNA ,Nucleic acid ,biology.protein ,artificial enzymes ,PF ,flavoenzyme mechanism ,Uridine Monophosphate ,Bacillus subtilis - Abstract
International audience; To facilitate production of functional enzymes and to study their mechanisms, especially in the complex cases of coenzyme-dependent systems, activation of an inactive apoenzyme preparation with a catalytically competent coenzyme intermediate is an attractive strategy. This is illustrated with the simple chemical synthesis of a flavin-methylene iminium compound previously proposed as a key intermediate in the catalytic cycle of several important flavoenzymes involved in nucleic acid metabolism. Reconstitution of both flavin-dependent RNA methyltransferase and thymidylate synthase apoproteins with this synthetic compound led to active enzymes for the C5-uracil methylation within their respective transfer RNA and dUMP substrate. This strategy is expected to be of general application in enzymology.
- Published
- 2017
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