Journal of Protein Chemistry

, Volume 16, Issue 5, pp 481–490 | Cite as

Peptide Sequencing by Mass Spectrometry for Homology Searches and Cloning of Genes

  • Andrej Shevchenko
  • Matthias Wilm
  • Matthias Mann
Article

Abstract

It is now possible to obtain sequence information from gel-separated proteins by mass spectrometry at levels too low for conventional approaches. Usually this tandem mass spectrometric data are used for database searches with the aim of identifying the corresponding gene. Recently it has been shown that long and accurate amino acid sequences can be obtained which are sufficient for PCR-based strategies to clone the corresponding gene [Wilm et al. (1996), Nature379, 466–469]. More than eight proteins have now been cloned based on that method. In many more cases the sequence information identified homologous proteins. Issues involved in cloning by mass spectrometric sequence information are discussed, as are two case studies. These results clearly establish mass spectrometry as a viable tool not only for the database identification of proteins, but also for the de novo sequencing of gel-separated proteins at the low-picomole to femtomole level.

Mass spectrometry protein sequencing electrospray protein identification 

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Copyright information

© Plenum Publishing Corporation 1997

Authors and Affiliations

  • Andrej Shevchenko
    • 1
  • Matthias Wilm
    • 1
  • Matthias Mann
    • 1
  1. 1.Protein & Peptide GroupEuropean Molecular Biology LaboratoryHeidelbergGermany

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