Skip to main content
Log in

Fragmentation of phosphopeptides in an ion trap mass spectrometer

  • Articles
  • Published:
Journal of the American Society for Mass Spectrometry

Abstract

A systematic study of the fragmentation pattern of phosphopeptides in an electrospray (ESI) ion trap mass spectrometer is presented. We show that phosphotyrosine- and phosphothreonine-containing peptides show complicated fragmentation patterns. These phosphopeptides were observed to lose the phosphate moiety in the form of H3PO4 and/or HPO3, but were also detected with no loss of the phosphate group. The tendency to lose the phosphate moiety depends strongly on the charge state. Thus, the highest observed charge state tends to retain the phosphate moiety with extensive fragmentation along the peptide backbone. We also show that phosphoserine-containing peptides have relatively simple fragmentation patterns of losing H3PO4. This loss is independent of the charge state. We suggest strategies for the accurate identification of phosphorylation sites using the ion trap mass spectrometer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Gibson, B. W.; Cohen, P. Methods of Enzymology, Volume 193: Mass Spectrometry; Academic: San Diego, CA, 1990; 480–501.

    Google Scholar 

  2. Bradshaw, R. A.; Stewart, A. E. Curr. Op. Biotech. 1994, 5, 85–93.

    Article  CAS  Google Scholar 

  3. Huddleston, M. J.; Annan, R. S.; Bean, M. F.; Carr, S. A. J. Am. Soc. Mass Spectrom. 1993, 4, 710–717.

    Article  CAS  Google Scholar 

  4. Nuwaysir, L. M.; Stults, J. T. J. Am. Soc. Mass Spectrom. 1993, 4, 662–669.

    Article  CAS  Google Scholar 

  5. Ding, J.; Burkhart, W.; Kassel, D. B. Rapid Commun. Mass Spectrom. 1994, 8, 94–98.

    Article  CAS  Google Scholar 

  6. Hunter, A. P.; Games, D. E. Rapid Commun. Mass Spectrom. 1994, 8, 559–570.

    Article  CAS  Google Scholar 

  7. Liao, P.; Leykam, J.; Andrews, P. C.; Gage, D. A.; Allison, J. Anal. Biochem. 1994, 219, 9–20.

    Article  CAS  Google Scholar 

  8. Annan, R. S.; Carr, S. A. Anal. Chem. 1996, 68, 3413–3421.

    Article  CAS  Google Scholar 

  9. Carr, S. A.; Huddleston, M. J.; Annan, R. S. Anal. Biochem. 1996, 239, 180–192.

    Article  CAS  Google Scholar 

  10. Jonscher, K. R.; Yates, J. R., III. J. Biol. Chem. 1997, 272, 1735–1741.

    Article  CAS  Google Scholar 

  11. Qin, J.; Chait, B. T. Anal. Chem. 1997, 69, 4002–4009.

    Article  CAS  Google Scholar 

  12. Tang, X. J.; Thibault, P.; Boyd, R. K. Anal. Chem. 1993, 65, 2824–2834.

    Article  CAS  Google Scholar 

  13. Zhang, X.; Herring, C. J.; Romano, P. R.; Szczepanowska, J.; Brzeska, H.; Hinnebusch, A. G.; Qin, J. Anal. Chem. 1998, 70, 2050–2059.

    Article  CAS  Google Scholar 

  14. Szczepanowska, J.; Zhang, X.; Herring, C. J.; Qin, J.; Korn, E. D.; Brzeska, H. Proc. Natl. Acad. Sci. USA 1997, 94, 8503–8508.

    Article  CAS  Google Scholar 

  15. Gillece-Castro, B. L.; Stults, J. T., Proceedings of the 43rd ASMS Conference on Mass Spectrometry and Allied Topics; Atlanta, GA, 1995; p 302.

  16. Haller, I.; Mirza, U. A.; Chait, B. T. Proceedings of the 45th ASMS Conference on Mass Spectrometry and Allied Topics; Atlanta, GA, 1997; p 611.

  17. Brzeska, H.; Szczepanowska, J.; Hoey, J.; Korn, E. D. J. Biol. Chem. 1996, 271, 27056–27062.

    Article  CAS  Google Scholar 

  18. Wilm, M.; Mann, M. Anal. Chem. 1996, 68, 1–8.

    Article  CAS  Google Scholar 

  19. Biemann, K. Biomed. Environ. Mass Spectrom. 1988, 16, 99–111.

    Article  CAS  Google Scholar 

  20. Sang-Won, L.; Kim, H. S.; Beauchamp, J. L. J. Am. Chem. Soc. 1998, 120, 3188–3195.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jun Qin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

DeGnore, J.P., Qin, J. Fragmentation of phosphopeptides in an ion trap mass spectrometer. J Am Soc Mass Spectrom 9, 1175–1188 (1998). https://doi.org/10.1016/S1044-0305(98)00088-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1016/S1044-0305(98)00088-9

Keywords

Navigation