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UDP-galactopyranose mutase has a novel structure and mechanism

Nature Structural Biology volume 8, pages 858–863 (2001)Cite this article

Abstract

Uridine diphosphogalactofuranose (UDP-Galf ) is the precursor of the d-galactofuranose (Galf ) residues found in bacterial and parasitic cell walls, including those of many pathogens, such as Mycobacterium tuberculosis and Trypanosoma cruzi. UDP-Galf is made from UDP-galactopyranose (UDP-Galp) by the enzyme UDP-galactopyranose mutase (mutase). The mutase enzyme is essential for the viability of mycobacteria and is not found in humans, making it a viable therapeutic target. The mechanism by which mutase achieves the unprecedented ring contraction of a nonreducing sugar is unclear. We have solved the crystal structure of Escherichia coli mutase to 2.4 Å resolution. The novel structure shows that the flavin nucleotide is located in a cleft lined with conserved residues. Site-directed mutagenesis studies indicate that this cleft contains the active site, with the sugar ring of the substrate UDP-galactose adjacent to the exposed isoalloxazine ring of FAD. Assay results establish that the enzyme is active only when flavin is reduced. We conclude that mutase most likely functions by transient reduction of substrate.

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Figure 1: Role of UDP-galactopyranose mutase in the biosynthesis of the cell wall.
Figure 2: Structure of UDP-galactopyranose mutase from E. coli.
Figure 3: Putative active site of UDP-galactopyranose mutase.

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Acknowledgements

The work is supported by the BBSRC. We thank W. Crocker, S. Chapman, S. Daff, R. Harris, N. Scrutton and G. Leonard for their help with experiments. We thank the reviewers and editor for assistance in revising the manuscript. We thank J. Blanchard and J. Barlow for helpful discussions. J.H.N. is a BBSRC Career Development Fellow. C.W. is a CIHR Senior Scientist and is supported by the Canadian Bacterial Diseases Network (NCE program).

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  1. Stephen A. McMahon

    Present address: Department of Chemistry, University of California at San Diego, 9500 Gilman Drive, La Jolla, California, 92093-0314, USA

Authors and Affiliations

  1. The Centre for Biomolecular Sciences, The University, St. Andrews, KY16 9ST, Scotland, UK

    David A.R. Sanders, Adam G. Staines, Stephen A. McMahon & James H. Naismith

  2. Department of Microbiology, Colorado State University, Fort Collins, 80523, Colorado, USA

    Michael R. McNeil

  3. Department of Microbiology, The University of Guelph, Ontario, N1G 2W1, Canada

    Chris Whitfield

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  1. David A.R. Sanders
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Correspondence to James H. Naismith.

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Sanders, D., Staines, A., McMahon, S. et al. UDP-galactopyranose mutase has a novel structure and mechanism. Nat Struct Mol Biol 8, 858–863 (2001). https://doi.org/10.1038/nsb1001-858

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