Skip to main content
Log in

The Essential Role of Mass Spectrometry in Characterizing Protein Structure: Mapping Posttranslational Modifications

  • Published:
Journal of Protein Chemistry Aims and scope Submit manuscript

Abstract

Over the last few years we have developed mass spectrometry-based approaches for selective identification of a variety of posttranslational modifications, and for sequencing the modified peptides. These methods do not involve radiolabeling or derivatization. Instead, modification-specific fragment ions are produced by collision-induced dissociation (CID) during analysis of peptides by ESMS. The formation and detection of these marker ions on-the-fly during the LC-ESMS analysis of a protein digest is a powerful technique for identifying posttranslationally modified peptides. Using the marker ion strategy in an orthogonal fashion, a precursor ion scan can detect peptides which give rise to a diagnostic fragment ion, even in an unfractionated protein digest. Once the modified peptide has been located, the appropriate precursor ion can be sequenced by tandem MS. The utility and interplay of this approach to mapping PTM is illustrated with examples that involve protein glycosylation and phosphorylation.

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

  • Annan, R. S., and Carr, S. A. (1996). Anal. Chem. 68, 3413–3421.

    Article  CAS  PubMed  Google Scholar 

  • Annan, R. S., McNulty, D. E., and Carr, S. A. (1996a). Proceedings 44th ASMS Conference on Mass Spectrometry and Allied Topics, Portland, Oregon, p. 702.

  • Annan, R. S., Huddleston, M. J., and Carr, S. A., (1996b). Proceedings 44th ASMS Conference on Mass Spectrometry and Allied Topics, Portland, Oregon, p. 1107.

  • Biemann, K. (1992). Mass spectrometry of peptides and proteins, Ann. Rev. Biochem. 61, 977–1010.

    Article  CAS  PubMed  Google Scholar 

  • Bradbury N. E., and Nielsen R. A., (1936). Phys. Rev. 49, 388–393.

    Article  CAS  Google Scholar 

  • Carr, S. A., Hemling, M. E., Bean, M. F., and Roberts, G. D. (1991). Anal. Chem. 63, 2802–2824.

    Article  CAS  PubMed  Google Scholar 

  • Carr, S. A., Huddleston, M. J., and Bean, M. F. (1993). Protein Sci. 2, 183–196.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Carr, S. A., Roberts, G., Annan, R. S., Hemling, M. E., and Hoyes, J. (1995). In Proceedings 43rd ASMS Conference on Mass Spectrometry and Allied Topics, Atlanta, Georgia, p. 620.

  • Carr, S. A., Huddleston, M. J., and Annan, R. S. (1996). Anal. Biochem. 239, 180–192.

    Article  CAS  PubMed  Google Scholar 

  • Chait, B. T., and Kent, S. B. H. (1992), Science 257, 1885–1894.

    Article  CAS  PubMed  Google Scholar 

  • Cohen, P., and Klee C., (1988) Calmodulin, Elsevier, New York.

    Google Scholar 

  • Denhardt, D. T., and Guo, X. (1993). FASEB J. 7, 1475–1482.

    Article  CAS  PubMed  Google Scholar 

  • Fenn, J. B., Mann, M., Meng, C. K., Wong, S. F., and Whitehouse, C. M. (1990). Mass Spectrom. Rev. 9, 37–70.

    Article  CAS  Google Scholar 

  • Hess, D., Covey, T. C., Winz, R., Brownsey, R. W., and Aebersold, R. (1993). Protein Sci. 2, 1342–1351.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hubbard, M. J., and Cohen, P. (1993). Trends Biochem. Sci. 18, 172–177.

    Article  CAS  PubMed  Google Scholar 

  • Huddleston, M. J., Annan, R. S., M. J., Bean, M. F., and Carr, S. A. (1993a). J. Am. Soc. Mass Spectrom. 4, 710–717.

    Article  CAS  PubMed  Google Scholar 

  • Huddleston, M. J., Bean, M. F., and Carr, S. A. (1993b). Anal. Chem. 66, 877–884

    Article  Google Scholar 

  • Kaufmann, R., Spengler, B., and Lutzenkirchen, F. (1993). Rapid Commun. Mass Spectrom. 7, 902–910.

    Article  CAS  PubMed  Google Scholar 

  • Thomson, B. A., Douglas, D. J., Corr, J. J., Hager, J. W., and Jollife, C. L. (1995). Anal. Chem., 67, 1696–1704.

    Article  CAS  Google Scholar 

  • Vestal, M. L., Juhasz, P., and Martin, S. A. (1995). Rapid Commun. Mass Spectrom. 9, 1044–1050.

    Article  CAS  Google Scholar 

  • Wilkins, M. R., Sanchez, J-C., Gooley, A. A., Appel, R. D., Humphery-Smith, I., Hochstrasser, D. F., and Williams, K. L. (1995). Biotechn. Gen. Eng. Rev. 13, 19–50.

    Article  Google Scholar 

  • Wilm, M., and Mann, M. (1994). Int. J. Mass Spectrom. Ion Process. 136, 167–180.

    Article  CAS  Google Scholar 

  • Wilm, M., and Mann, M. (1996) Anal. Chem. 68, 1–8.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Annan, R.S., Carr, S.A. The Essential Role of Mass Spectrometry in Characterizing Protein Structure: Mapping Posttranslational Modifications. J Protein Chem 16, 391–402 (1997). https://doi.org/10.1023/A:1026384605285

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1026384605285

Navigation