Mass Spectrometry: Peptide Sequencing

J Throck Watson, Michigan State University, East Lansing, Michigan, USA

Published online: April 2001

DOI: 10.1038/npg.els.0003111

Electrospray and MALDI (matrix-assisted laser desorption ionization) are two versions of desorption/ionization that allow modern mass spectrometry to be applied to the analysis of peptides and proteins. In addition to a direct indication of the molecular weight, the sequence of amino acids in a peptide can be deduced from the fragmentation pattern (the mass spectrum itself).

Keywords: electrospray; MALDI; signal suppression; mass spectrum; matrix-assisted laser desorption ionization

Figure 1. Conceptual respresentation of key bonds in a given peptide that break to form the indicated N-terminal (a, b, c) or C-terminal (x, y, z) fragments.
Figure 2. ES-CAD-MS/MS mass spectrum of a peptide, LKRApTLG-amide.
Figure 3. ES-CAD-MS/MS mass spectrum of Ac-TMPP derivative of LKRApTLG-amide. The charged derivative (Ac-TMPP) of the peptide promotes remote-site fragmentation, which gives a more complete series of fragment ions (compared to that in Figure 2) from which the sequence and locus of the phosphate group can be deduced.
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 References
    Cotter R (1992) MALDI-TOF-MS. Analytical Chemistry 64: 1027A–1039A.
    Hofstadler SA, Bakhtiar R and Smith RD (1996) Instrumentation and spectral interpretation for electrospray MS. Journal of Chemical Education 73: A82–A88.
    Johnson RS, Martin SA, Biemann K, Stults JT and Watson JT (1987) Novel fragmentation process of peptides by CAD-MS/MS: differentiation of leucine and isoleucine. Analytical Chemistry 59: 2621–2625.
    Kolli VSK and Orlando R (1995) Complete sequence confirmation of large peptides by high energy CAD. Journal of the American Society of Mass Spectrometry 6: 234–241.
    Roepstorff P and Fohlman J (1984) Nomenclature for MS of peptides. Biomedical Mass Spectrometry 11: 601.
    Shen T-L, Huang Z-H, Laivenieks M et al. (1999) Evaluation of charge derivatization of a proteolytic digest for improved mass spectrometric analysis. Journal of Mass Spectrometry 34: 1154–1165.
    Yates JR, Eng JK and McCormack AL (1995) Correlating MS/MS of peptides to sequences in nucleotide databases. Analytical Chemistry 67: 3202–3210.
 Further Reading
    book Watson JT (1997) Introduction to Mass Spectrometry, 3rd edn. Philadelphia: Lippincott-Raven.

Related Sub-Topics:

Protein Structure | Mass Spectrometry | Biochemical and Biophysical Techniques | Proteins: Structure, Function, Metabolism
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Article Title: Mass Spectrometry: Peptide Sequencing
 
How to Cite close
Watson, J Throck(Apr 2001) Mass Spectrometry: Peptide Sequencing. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003111]