Brief Communication | Published:

Legionella effector SetA as a general O-glucosyltransferase for eukaryotic proteins

Nature Chemical Biologyvolume 15pages213216 (2019) | Download Citation

Abstract

The identification of host protein substrates is key to understanding effector glycosyltransferases secreted by pathogenic bacteria and to using them for glycoprotein engineering. Here we report a chemical method for tagging, enrichment, and site-specific proteomic profiling of effector-modified proteins in host cells. Using this method, we discover that Legionella effector SetA α-O-glucosylates various eukaryotic proteins by recognizing a S/T-X-L-P/G sequence motif, which can be exploited to site-specifically introduce O-glucose on recombinant proteins.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Data availability

The data that support the findings of this study are available from the corresponding author on reasonable request.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  1. 1.

    Schmaltz, R. M., Hanson, S. R. & Wong, C.-H. Chem. Rev. 111, 4259–4307 (2011).

  2. 2.

    Moremen, K. W. et al. Nat. Chem. Biol. 14, 156–162 (2017).

  3. 3.

    Nothaft, H. & Szymanski, C. M. Nat. Rev. Microbiol. 8, 765–778 (2010).

  4. 4.

    Keys, T. G. & Aebi, M. Curr. Opin. Syst. Biol. 5, 23–31 (2017).

  5. 5.

    Schwarz, F. et al. Nat. Chem. Biol. 6, 264–266 (2010).

  6. 6.

    Valderrama-Rincon, J. D. et al. Nat. Chem. Biol. 8, 434–436 (2012).

  7. 7.

    Naegeli, A. et al. J. Biol. Chem. 289, 24521–24532 (2014).

  8. 8.

    Xu, Y. et al. Chem. Commun. (Camb.) 53, 9075–9077 (2017).

  9. 9.

    Kightlinger, W. et al. Nat. Chem. Biol. 14, 627–635 (2018).

  10. 10.

    Lu, Q., Li, S. & Shao, F. Trends Microbiol. 23, 630–641 (2015).

  11. 11.

    Jank, T., Belyi, Y. & Aktories, K. Cell Microbiol. 17, 1752–1765 (2015).

  12. 12.

    Sun, Y., Willis, L. M., Batchelder, H. R. & Nitz, M. Chem. Commun. (Camb.) 52, 13024–13026 (2016).

  13. 13.

    Just, I. et al. Nature 375, 500–503 (1995).

  14. 14.

    Belyi, Y. et al. Proc. Natl. Acad. Sci. USA 103, 16953–16958 (2006).

  15. 15.

    López Aguilar, A. et al. ACS. Chem. Biol. 12, 611–621 (2017).

  16. 16.

    Jank, T. et al. Cell Microbiol. 14, 852–868 (2012).

  17. 17.

    O’Shea, J. P. et al. Nat. Methods 10, 1211–1212 (2013).

  18. 18.

    Rana, N. A. & Haltiwanger, R. S. Curr. Opin. Struct. Biol. 21, 583–589 (2011).

  19. 19.

    Yu, H. et al. Nat. Chem. Biol. 12, 735–740 (2016).

  20. 20.

    Steinemann, M., Schlosser, A., Jank, T. & Aktories, K. Proc. Natl. Acad. Sci. USA 115, 9580–9585 (2018).

  21. 21.

    Levanova, N. et al. Naunyn Schmiedebergs Arch. Pharmacol. 322, 390 (2018).

  22. 22.

    Wang, Z. et al. Cell Disc. 4, 53 (2018).

  23. 23.

    Shen, D. L. et al. ACS Chem. Biol. 12, 206–213 (2017).

  24. 24.

    Darabedian, N., Gao, J., Chuh, K. N., Woo, C. M. & Pratt, M. R. J. Am. Chem. Soc. 140, 7092–7100 (2018).

  25. 25.

    Li, S. et al. Nature 501, 242–246 (2013).

  26. 26.

    Berger, K. H. & Isberg, R. R. Mol. Microbiol. 7, 7–19 (1993).

  27. 27.

    Besanceney-Webler, C. et al. Angew. Chem. Int. Ed. Engl. 50, 8051–8056 (2011).

  28. 28.

    Qin, K. et al. ACS Chem. Biol. 13, 1983–1989 (2018).

  29. 29.

    Xu, L. et al. PLoS Pathog. 6, e1000822 (2010).

  30. 30.

    Crooks, G. E., Hon, G., Chandonia, J.-M. & Brenner, S. E. Genome Res. 14, 1188–1190 (2004).

  31. 31.

    Eswar, N. et al. Curr. Protoc. Bioinformatics Chapter 5, Unit 5.6 (2006).

  32. 32.

    Larkin, M. A. et al. Bioinformatics 23, 2947–2948 (2007).

  33. 33.

    Maier, J. A. et al. J. Chem. Theory. Comput. 11, 3696–3713 (2015).

  34. 34.

    Wang, J., Wolf, R. M., Caldwell, J. W., Kollman, P. A. & Case, D. A. J. Comput. Chem. 25, 1157–1174 (2004).

  35. 35.

    Petersen, H. G. J. Chem. Phys. 103, 3668–3679 (1995).

Download references

Acknowledgements

We thank Z. Luo for providing L. pneumophila strains, W. Zhou at the mass spectrometry facility of the National Center for Protein Sciences at Peking University for assistance with proteomics analysis, and the High Performace Computing Platform of the Center for Life Sciences for supporting protein structure modeling. This work is supported by the National Key R&D Program of China (numbers 2018YFA0507600 and 2016YFA0501500 to X.C. and 2016YFA0502300 to L.L.) and the National Natural Science Foundation of China (numbers 21425204, 21672013, 91753206, and 21521003 to X.C.).

Author information

Affiliations

  1. College of Chemistry and Molecular Engineering, Peking University, Beijing, China

    • Ling Gao
    • , Qitao Song
    • , Hao Liang
    • , Yuntao Zhu
    • , Luhua Lai
    •  & Xing Chen
  2. Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China

    • Ling Gao
    • , Qitao Song
    • , Hao Liang
    • , Tiantian Wei
    • , Junyu Xiao
    • , Luhua Lai
    •  & Xing Chen
  3. Beijing National Laboratory for Molecular Sciences, Peking University, Beijing, China

    • Yuntao Zhu
    • , Luhua Lai
    •  & Xing Chen
  4. School of Life Sciences, Peking University, Beijing, China

    • Tiantian Wei
    •  & Junyu Xiao
  5. State Key Lab of Animal Nutrition, Ministry of Agriculture Feed Industry Center, China Agricultural University, Beijing, China

    • Na Dong
  6. National Institute of Biological Sciences, Beijing, China

    • Na Dong
    •  & Feng Shao
  7. Center for Quantitative Biology, Peking University, Beijing, China

    • Luhua Lai
  8. Synthetic and Functional Biomolecules Center, Peking University, Beijing, China

    • Xing Chen
  9. Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Peking University, Beijing, China

    • Xing Chen

Authors

  1. Search for Ling Gao in:

  2. Search for Qitao Song in:

  3. Search for Hao Liang in:

  4. Search for Yuntao Zhu in:

  5. Search for Tiantian Wei in:

  6. Search for Na Dong in:

  7. Search for Junyu Xiao in:

  8. Search for Feng Shao in:

  9. Search for Luhua Lai in:

  10. Search for Xing Chen in:

Contributions

L.G. and X.C. conceived the project; L.G. performed experiments with the help of Q.S., Y.Z., T.W., N.D., J.X., and F.S.; H.L. and L.L. performed the structural modeling; L.G. and X.C. analyzed the data and wrote the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Xing Chen.

Supplementary information

  1. Supplementary Text and Figures

    Supplementary Figures 1–16, Supplementary Note 1

  2. Reporting Summary

  3. Supplementary Dataset 1

About this article

Publication history

Received

Accepted

Published

Issue Date

DOI

https://doi.org/10.1038/s41589-018-0189-y