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Research article2011Peer reviewedOpen access

Structural and functional studies of mycobacterial IspD enzymes

Björkelid, Christofer; Bergfors, Terese; Henriksson, Lena M.; Stern, Ana Laura; Mowbray, Sherry; Unge, Torsten; Jones, T. Alwyn

Abstract

A number of pathogens, including the causative agents of tuberculosis and malaria, synthesize isopentenyl diphosphate via the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway rather than the classical mevalonate pathway found in humans. As part of a structure-based drug-discovery program against tuberculosis, IspD, the enzyme that carries out the third step in the MEP pathway, was targeted. Constructs of both the Mycobacterium smegmatis and the Mycobacterium tuberculosis enzymes that were suitable for structural and inhibitor-screening studies were engineered. Two crystal structures of the M. smegmatis enzyme were produced, one in complex with CTP and the other in complex with CMP. In addition, the M. tuberculosis enzyme was crystallized in complex with CTP. Here, the structure determination and crystallographic refinement of these crystal forms and the enzymatic characterization of the M. tuberculosis enzyme construct are reported. A comparison with known IspD structures allowed the definition of the structurally conserved core of the enzyme. It indicates potential flexibility in the enzyme and in particular in areas close to the active site. These well behaved constructs provide tools for future target-based screening of potential inhibitors. The conserved nature of the extended active site suggests that any new inhibitor will potentially exhibit broad-spectrum activity.

Published in

Acta Crystallographica Section D: Biological Crystallography
2011, Volume: 67, pages: 403-414
Publisher: WILEY-BLACKWELL

      SLU Authors

    • Stern, Ana Laura

      • Department of Molecular Biology, Swedish University of Agricultural Sciences

      • Mowbray, Sherry

        • Department of Molecular Biology, Swedish University of Agricultural Sciences

      Sustainable Development Goals

      Ensure healthy lives and promote well-being for all at all ages

      UKÄ Subject classification

      Biochemistry and Molecular Biology

      Publication identifier

      DOI: https://doi.org/10.1107/S0907444911006160

      Permanent link to this page (URI)

      https://res.slu.se/id/publ/46733