Abstract
MALDI-TOF peptide mass fingerprinting (PMF) is the fastest and cheapest method of protein identification; the studied genome is sequenced and annotated, and the protein is amenable to separation and detection in 2D gel electrophoresis. In plant proteomics there are two main difficulties: few plant genomes are sequenced, and major contaminants are non-plant specific. This chapter describes the classical “bottom-up” method (i.e., from peptide to protein identification) of gel cutting, in-gel digestion, peptide recovery and purification, MALDI-TOF mass spectrometry, and critical survey of protein database queries.
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References
Karas M., Bachman D., Bahr U., and Hillenkamp F. (1987) Matrix-assisted ultraviolet laser desorption of non-volatile compounds. Int. J. Mass Spectrom. Ion Proc. 53–68.
Karas M. and Hillenkamp F. (1988) Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons. Anal. Chem. 2299–2301.
Karas M., Bahr U., Ingendoh A., and Hillenkamp F. (1989) Laser desorption/ionisation mass spectrometry of proteins of mass 100,000 to 250,000 Dalton. Angewandte Chemie Int. Ed. 760–761.
Fenn J. B., Mann M., Meng C. K., Wong S. F., and Whitehouse C. M. (1989) Electrospray ionization for mass spectrometry of large biomolecules. Science 64–71.
http://www.nobelprize.org/chemistry/laureates/2002/index.html.
Tanaka K., Waki H., Ido Y., Akita S., Yoshida Y., and Yoshida T. (1988) Protein and polymer analysis up to m/z 100,000 by laser ionisation time-of-flight mass spectrometry. Rapid Comm. Mass Spectrom. 151–153.
Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plant. Nature 796–815.
Jensen O., Wilm M., Schevschenko A., and Mann M. (1999) Sample preparation methods for mass spectrometric peptide mapping directly from 2-DE gels, in Methods in Molecular Biology: 2-D Proteome Analysis Protocols, vol. 112 (Link, A. J., ed.), Humana, Totowa, NJ, pp. 513–530.
De Hoffmann, E., Charette, J., and Stroobant, V. (1999) Spectrométrie de Masse, Cours et Exercices Corrigés, 2ème édition. Dunod, Paris.
De Hoffmann, E. and Stroobant, V. (2002) Mass Spectrometry: Principles and Applications, 2nd ed. John Wiley & Sons, Hoboken, NJ.
Throck Watson, J. (1997) Introduction to Mass Spectrometry, 3rd ed. Lippincott-Raven: New York.
James, P. (ed.) (2000) Proteome Research: Mass Spectrometry (Principles and Practice). Springer Verlag, New York.
Cotter, R. J. (1997) Time-of-Flight Mass Spectrometry: Instrumentation and Applications in Biological Research. American Chemical Society, Washington, DC.
Vestal M. L., Juhasz P., Hines W., and Martin S. (1998) in Proceedings of the 46th ASMS Conference on Mass Spectrometry and Allied Topics, Orlando, FL.
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Sommerer, N., Centeno, D., Rossignol, M. (2007). Peptide Mass Fingerprinting. In: Thiellement, H., Zivy, M., Damerval, C., Méchin, V. (eds) Plant Proteomics. Methods in Molecular Biology, vol 355. Humana Press. https://doi.org/10.1385/1-59745-227-0:219
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DOI: https://doi.org/10.1385/1-59745-227-0:219
Publisher Name: Humana Press
Print ISBN: 978-1-58829-635-1
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