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Research article2010Peer reviewed

Crystal Structures of an Oligopeptide-Binding Protein from the Biosynthetic Pathway of the beta-Lactamase Inhibitor Clavulanic Acid

Mackenzie Alasdair K., Valegard Karin, Iqbal Aman, Caines Matthew E. C., Kershaw Nadia J., Jensen Susan E., Schofield Christopher J., Andersson Inger

Abstract

Clavulanic acid (CA) is a clinically important ß-lactamase inhibitor that is produced by fermentation of Streptomyces clavuligerus. The CA biosynthesis pathway starts from arginine and glyceraldehyde-3-phosphate, and proceeds via (3S,5S)-clavaminic acid, which is converted to (3R,5R)-clavaldehyde, the immediate precursor of (3R,5R)-CA. Open reading frames 7 (orf7) and 15 (orf15) of the CA biosynthesis cluster encode oligopeptide binding proteins (OppA1 and OppA2), that are essential for CA biosynthesis. OppA1/2 are proposed to be involved in the binding and/or transport of peptides across the S. clavuligerus cell membrane. Peptide binding assays reveal that recombinant OppA1 and 2 bind di/tri peptides containing arginine. Crystal structures of OppA2 in its apo form and in complex with arginine or bradykinin were solved to 1.45, 1.7, and 1.7 Å resolution, respectively. The overall fold of OppA2 consists of two lobes with a deep cavity in the centre, as observed for other oligopeptide binding proteins. The large cavity creates a peptide/arginine binding cleft. The crystal structures of OppA2 in complex with arginine or bradykinin reveal that the C-terminal arginine of bradykinin binds similarly to arginine. The results are discussed in terms of the possible roles of OppA1/2 in CA biosynthesis

Keywords

clavulanic acid; beta-lactam; beta-lactamase inhibitor; oligopeptide-binding proteins; solute-binding proteins

Published in

Journal of Molecular Biology
2010, Volume: 396, number: 2, pages: 332-344
Publisher: Elsevier

      SLU Authors

    • Mackenzie, Alasdair

      • Department of Molecular Biology, Swedish University of Agricultural Sciences

      • Valegård, Karin

        • Department of Molecular Biology, Swedish University of Agricultural Sciences

        • Andersson, Inger

          • Department of Molecular Biology, Swedish University of Agricultural Sciences

        UKÄ Subject classification

        Biochemistry and Molecular Biology

        Publication identifier

        DOI: https://doi.org/10.1016/j.jmb.2009.11.045

        Permanent link to this page (URI)

        https://res.slu.se/id/publ/28924