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Secondary structure reshuffling modulates glycosyltransferase function at the membrane

Nature Chemical Biology volume 11, pages 16–18 (2015)Cite this article

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Abstract

Secondary structure refolding is a key event in biology as it modulates the conformation of many proteins in the cell, generating functional or aberrant states. The crystal structures of mannosyltransferase PimA reveal an exceptional flexibility of the protein along the catalytic cycle, including β-strand–to–α-helix and α-helix–to–β-strand transitions. These structural changes modulate catalysis and are promoted by interactions of the protein with anionic phospholipids in the membrane.

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Figure 1: Overall structure of PimA in the absence of ligands.
Figure 2: The canonical GT-B conformation (GDP-bound form) of PimA, but not the atypical apo form, is functionally competent for nucleotide-sugar binding and catalysis.
Figure 3: The N-terminal domain of PimA mediates protein-membrane association.

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Acknowledgements

We acknowledge A. Haouz and P. Weber (Institut Pasteur, France) for help with robotic crystallization; S. Lopez Fernandez and P. Arrasate (Structural Glycobiology Group, Unit of Biophysics, Spain) for technical assistance; and the European Synchrotron Radiation Facility (ESRF), the French National Synchrotron SOLEIL, the Diamond Light Source (DLS) and the Swiss Light Source (SLS) for granting access to synchrotron radiation facilities and their staff for the onsite assistance. We specially thank the BioStruct-X project to support access to structural biology facilities. Technical support provided by Universidad del País Vasco/Euskal Herriko Unibertsitatea and Ministerio de Ciencia e Innovación is acknowledged. We also thank all members of the Structural Glycobiology Group for valuable scientific discussions. This work was supported by the European Community's Sixth and Seventh Framework Programmes (contracts LSHP-CT-2005-018923 and HEALTH-F3-2011-260872), the Institut Pasteur, the Spanish Ministry of Science and Innovation (contracts SAF2010-19096 and BIO2013-49022-C2-2-R), IKERBASQUE, the Basque Foundation for Science, the Basque Government and the Fundación Biofísica Bizkaia.

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Authors and Affiliations

  1. Institut Pasteur, Unité de Microbiologie Structurale, CNRS UMR 3528, Paris, France

    David Giganti, Mariano A Martínez, Nathalie Barilone, Marco Bellinzoni & Pedro M Alzari

  2. Unidad de Biofisica, Centro Mixto Consejo Superior de Investigaciones Cientificas–Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Leioa, Bizkaia, Spain.,

    David Giganti, David Albesa-Jové, Saioa Urresti, Ane Rodrigo-Unzueta, Natalia Comino & Marcelo E Guerin

  3. Departamento de Bioquímica, Universidad del País Vasco, Spain

    David Giganti, David Albesa-Jové, Saioa Urresti, Ane Rodrigo-Unzueta, Natalia Comino & Marcelo E Guerin

  4. Institut Pasteur, Unité de Biochimie des Interactions Macromoléculaires, CNRS UMR 3528, Paris, France

    Alexandre Chenal

  5. IKERBASQUE, Basque Foundation for Science, Bilbao, Spain

    Marcelo E Guerin

Authors
  1. David Giganti
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Contributions

M.E.G. and P.M.A. conceived the project. D.G., D.A.-J. and M.B. performed the crystallographic studies. S.U., A.R.-U., M.A.M., N.B. and N.C. prepared and characterized wild-type PimA and the different mutants. D.G., M.A.M., M.B., A.C., M.E.G. & P.M.A. analyzed the results. D.G., A.C., M.E.G. & P.M.A. wrote the manuscript.

Corresponding authors

Correspondence to Marcelo E Guerin or Pedro M Alzari.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Figures 1–15 and Supplementary Table 1. (PDF 2845 kb)

Supplementary Video 1

Structural transition between the extended and compact conformational states of apo PimA (MPG 522 kb)

Supplementary Video 2

Structural transition between apo PimA and GDP-bound PimA involving the reshuffling of secondary structural elements from the N-terminal domain. (MPG 1520 kb)

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Giganti, D., Albesa-Jové, D., Urresti, S. et al. Secondary structure reshuffling modulates glycosyltransferase function at the membrane. Nat Chem Biol 11, 16–18 (2015). https://doi.org/10.1038/nchembio.1694

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