Skip to main content

N-Terminal Protein Ladder Sequencing

  • Protocol
Book cover Protein Sequencing Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 64))

  • 1001 Accesses

Abstract

In protein ladder sequencing, the amino acid sequence of a peptide is determined by measuring the molecular mass differences between members of a family of fragments produced from the peptide. Adjacent members of this family of sequence-defining peptide fragments differ from one another by one amino acid residue. The molecular masses of the family of peptide fragments are measured by mass spectrometry in a single operation to read-out of the amino acid sequence. The identity of the amino acid residues is assigned by the mass differences between adjacent peaks and the amino acid sequence is determined by the order of their occurrence in the data set.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Chart, B. T., Wang, R., Beavis, R. C., and Kent, S. B. H. (1993) Protein ladder sequencing. Science 262, 89–92.

    Article  Google Scholar 

  2. Wang, R., Chait, B. T., and Kent, S. B. H. (1994) Protein ladder sequencing towards automation, in Techniques in Protein Chemistry, vol. 5 (Crabb, J. W., ed.), Academic, San Diego, CA, pp. 19–26.

    Google Scholar 

  3. Bartlet-Jones, M., Jeffery, W. A., Hansen, H. F., and Pappin, D. J. C. (1994) Peptide ladder sequencing by mass spectrometry using a novel, volatile degradation reagent. Rapid Commun. Mass Spectrom 8, 737–742.

    Article  CAS  Google Scholar 

  4. Hillenkamp, F., Karas, M., Beavis, R. C., and Chait, B. T. (1991) Matrix-assisted laser desorption/ionization mass spectrometry of biopolymers. Anal. Chem. 63, 1193A–1203A.

    CAS  Google Scholar 

  5. Wang, R. and Chart, B. T. (1996) Post-translational modifications analyzed by automated protein ladder sequencing, in Protein and Peptide Analysis by Mass Spectrometry (Chapman, J. R., ed.), Humana, Totowa, NJ (in press).

    Google Scholar 

  6. Beavis, R. C. and Chait, B. T. (1989) Matrix-assisted laser-desorption mass spectrometry using 355 nm radiation. Rapid Commun. Mass Spectrom 3, 436–439.

    Article  CAS  Google Scholar 

  7. Tarr, G. E. (1977) Improved manual sequencing methods. Methods Enzymol 47, 335–357.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1997 Humana Press Inc.

About this protocol

Cite this protocol

Wang, R., Chait, B.T. (1997). N-Terminal Protein Ladder Sequencing. In: Smith, B.J. (eds) Protein Sequencing Protocols. Methods in Molecular Biology™, vol 64. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-353-8:175

Download citation

  • DOI: https://doi.org/10.1385/0-89603-353-8:175

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-353-5

  • Online ISBN: 978-1-59259-550-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics