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Research article2003Peer reviewedOpen access

Structure of Rhodococcus erythropolis limonene-1,2-epoxide hydrolase reveals a novel active site

Arand M, Hallberg BM, Zou JY, Bergfors T, Oesch F, van der Werf MJ, de Bont JAM, Jones TA, Mowbray SL

Abstract

Epoxide hydrolases are essential for the processing of epoxide-containing compounds in detoxification or metabolism. The classic epoxide hydrolases have an alpha/beta 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 Angstrom resolution. This enzyme represents a completely different structure and a novel one-step mechanism. The fold features a highly curved six-stranded mixed beta-sheet, with four alpha-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 Angstrom 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

Published in

EMBO Journal
2003, Volume: 22, number: 11, pages: 2583-2592
Publisher: OXFORD UNIV PRESS

      SLU Authors

    • Mowbray, Sherry

      • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

    UKÄ Subject classification

    Environmental Sciences related to Agriculture and Land-use

    Publication identifier

    DOI: https://doi.org/10.1093/emboj/cdg275

    Permanent link to this page (URI)

    https://res.slu.se/id/publ/890