Print Email Facebook Twitter Structure of Rhodococcus erythropolis limonene-1,2-epoxide hydrolase reveals a novel active site Title Structure of Rhodococcus erythropolis limonene-1,2-epoxide hydrolase reveals a novel active site Author Arand, M. Hallberg, B.M. Zou, J. Bergfors, T. Oesch, F. van der Werf, M.J. de Bont, J.A.M. Jones, T.A. Mowbray, S.L. Publication year 2003 Abstract Epoxide hydrolases are essential for the processing of epoxide-containing compounds in detoxification or metabolism. The classic epoxide hydrolases have an α/β hydrolase fold and act via a two-step reaction mechanism including an enzyme-substrate intermediate. We report here the structure of the limonene-1,2-epoxide hydrolase from Rhodococcus erythropolis, solved using single-wavelength anomalous dispersion from a selenomethionine-substituted protein and refined at 1.2 Å resolution. This enzyme represents a completely different structure and a novel one-step mechanism. The fold features a highly curved six-stranded mixed β-sheet, with four α-helices packed onto it to create a deep pocket. Although most residues lining this pocket are hydrophobic, a cluster of polar groups, including an Asp-Arg-Asp triad, interact at its deepest point. Site-directed mutagenesis supports the conclusion that this is the active site. Further, a 1.7 Å resolution structure shows the inhibitor valpromide bound at this position, with its polar atoms interacting directly with the residues of the triad. We suggest that several bacterial proteins of currently unknown function will share this structure and, in some cases, catalytic properties. Subject BiologyBiotechnologyCrystal structureEnantioselectivityEpoxide hydrolaseMechanismMonoterpene degradationbacterial enzymelimonene 1,2 epoxide hydrolaseselenomethionineunclassified drugvalpromidealpha helixarticlebeta sheetcatalysischemical reactioncrystal structuredetoxificationdrug protein bindingenantioselectivityenzyme active siteenzyme metabolismenzyme structureenzyme substrate complexnonhumanpriority journalreaction analysisRhodococcus erythropolissite directed mutagenesisAmino Acid SequenceBacterial ProteinsCatalytic DomainCrystallography, X-RayDimerizationEpoxide HydrolasesModels, MolecularMolecular Sequence DataMutagenesis, Site-DirectedProtein SubunitsRecombinant ProteinsRhodococcusSequence Homology, Amino AcidActinobacteria (class)Bacteria (microorganisms)RhodococcusRhodococcus erythropolisuncultured actinomycete To reference this document use: http://resolver.tudelft.nl/uuid:f69dfc5b-ab3e-4e1b-b31c-a44748e7e05a DOI https://doi.org/10.1093/emboj/cdg275 TNO identifier 237138 ISSN 0261-4189 Source EMBO Journal, 22 (11), 2583-2592 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.