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Structural snapshots of the reaction coordinate for O-GlcNAc transferase

Nature Chemical Biology volume 8, pages 966–968 (2012)Cite this article

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Abstract

Visualization of the reaction coordinate undertaken by glycosyltransferases has remained elusive but is critical for understanding this important class of enzyme. Using substrates and substrate mimics, we describe structural snapshots of all species along the kinetic pathway for human O-linked β-N-acetylglucosamine transferase (O-GlcNAc transferase), an intracellular enzyme that catalyzes installation of a dynamic post-translational modification. The structures reveal key features of the mechanism and show that substrate participation is important during catalysis.

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Figure 1: Structural snapshots define the kinetic mechanism of OGT.
Figure 2: Glycosylation reaction trajectory and interactions that facilitate glycosyl transfer.

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References

  1. Lairson, L.L., Henrissat, B., Davies, G.J. & Withers, S.G. Annu. Rev. Biochem. 77, 521–555 (2008).

    Article  CAS  Google Scholar 

  2. Torres, C.R. & Hart, G.W. J. Biol. Chem. 259, 3308–3317 (1984).

    CAS  PubMed  Google Scholar 

  3. Kreppel, L.K., Blomberg, M.A. & Hart, G.W. J. Biol. Chem. 272, 9308–9315 (1997).

    Article  CAS  Google Scholar 

  4. Sinclair, D.A. et al. Proc. Natl. Acad. Sci. USA 106, 13427–13432 (2009).

    Article  CAS  Google Scholar 

  5. Love, D.C. et al. Proc. Natl. Acad. Sci. USA 107, 7413–7418 (2010).

    Article  CAS  Google Scholar 

  6. Kazemi, Z., Chang, H., Haserodt, S., McKen, C. & Zachara, N.E. J. Biol. Chem. 285, 39096–39107 (2010).

    Article  CAS  Google Scholar 

  7. Dentin, R., Hedrick, S., Xie, J., Yates, J. III & Montminy, M. Science 319, 1402–1405 (2008).

    Article  CAS  Google Scholar 

  8. Lazarus, M.B., Nam, Y., Jiang, J., Sliz, P. & Walker, S. Nature 469, 564–567 (2011).

    Article  CAS  Google Scholar 

  9. Gloster, T.M. et al. Nat. Chem. Biol. 7, 174–181 (2011).

    Article  CAS  Google Scholar 

  10. Iyer, S.P. & Hart, G.W. J. Biol. Chem. 278, 24608–24616 (2003).

    Article  CAS  Google Scholar 

  11. Gross, B.J., Kraybill, B.C. & Walker, S. J. Am. Chem. Soc. 127, 14588–14589 (2005).

    Article  CAS  Google Scholar 

  12. Lee, S.S. et al. Nat. Chem. Biol. 7, 631–638 (2011).

    Article  CAS  Google Scholar 

  13. Burkart, M.D. et al. Bioorg. Med. Chem. 8, 1937–1946 (2000).

    Article  CAS  Google Scholar 

  14. Vocadlo, D.J. & Davies, G.J. Curr. Opin. Chem. Biol. 12, 539–555 (2008).

    Article  CAS  Google Scholar 

  15. Schramm, V.L. & Shi, W. Curr. Opin. Struct. Biol. 11, 657–665 (2001).

    Article  CAS  Google Scholar 

  16. Vocadlo, D.J., Davies, G.J., Laine, R. & Withers, S.G. Nature 412, 835–838 (2001).

    Article  CAS  Google Scholar 

  17. Davies, G.J., Planas, A. & Rovira, C. Acc. Chem. Res. 45, 308–316 (2012).

    Article  CAS  Google Scholar 

  18. Martinez-Fleites, C. et al. Nat. Struct. Mol. Biol. 15, 764–765 (2008).

    Article  CAS  Google Scholar 

  19. Dorfmueller, H.C. et al. Amino Acids 40, 781–792 (2011).

    Article  CAS  Google Scholar 

  20. Errey, J.C. et al. Angew. Chem. Int. Edn Engl. 49, 1234–1237 (2010).

    Article  CAS  Google Scholar 

  21. Reinert, D.J., Jank, T., Aktories, K. & Schulz, G.E. J. Mol. Biol. 351, 973–981 (2005).

    Article  CAS  Google Scholar 

  22. Shen, D.L., Gloster, T.M., Yuzwa, S.A. & Vocadlo, D.J. J. Biol. Chem. 287, 15395–15408 (2012).

    Article  CAS  Google Scholar 

  23. Ramakrishnan, B., Ramasamy, V. & Qasba, P.K. J. Mol. Biol. 357, 1619–1633 (2006).

    Article  CAS  Google Scholar 

  24. Persson, K. et al. Nat. Struct. Biol. 8, 166–175 (2001).

    Article  CAS  Google Scholar 

  25. Ni, L. et al. Biochemistry 46, 6288–6298 (2007).

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the US National Institutes of Health (NIH) grant R01GM094263 to S.W. and the Natural Sciences and Engineering Research Council of Canada and Simon Fraser University. T.M.G. is a Sir Henry Wellcome postdoctoral fellow and a Michael Smith for Health Research (MSFHR) trainee award holder. D.J.V. is an E.W.R. Steacie Memorial Fellow and holds a Canada Research Chair in Chemical Glycobiology. We thank L. Deng for assistance with the chemical synthesis. We thank L. Cai (University of South Carolina, Salkehatchie) for the kind gift of UDP-2-ketoGlc.

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Author notes

  1. Michael B Lazarus and Jiaoyang Jiang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA

    Michael B Lazarus

  2. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA

    Michael B Lazarus, Jiaoyang Jiang & Suzanne Walker

  3. Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada

    Tracey M Gloster, Wesley F Zandberg, Garrett E Whitworth & David J Vocadlo

  4. Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia, Canada

    David J Vocadlo

Authors
  1. Michael B Lazarus
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  2. Jiaoyang Jiang
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  3. Tracey M Gloster
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  4. Wesley F Zandberg
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  5. Garrett E Whitworth
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  7. Suzanne Walker
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Contributions

M.B.L. carried out all structural experiments. J.J. helped with X-ray data collection. J.J. and M.B.L. performed kinetics assays. W.F.Z. and T.M.G. performed thiosugar kinetics assays and peptide assays. G.E.W. performed the synthesis of peptide and glycopeptides substrates. M.B.L., J.J., T.M.G., D.J.V. and S.W. designed experiments, analyzed data and wrote the manuscript.

Corresponding author

Correspondence to Suzanne Walker.

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The authors declare no competing financial interests.

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Lazarus, M., Jiang, J., Gloster, T. et al. Structural snapshots of the reaction coordinate for O-GlcNAc transferase. Nat Chem Biol 8, 966–968 (2012). https://doi.org/10.1038/nchembio.1109

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