Two-dimensional reference map of Agrobacterium tumefaciens proteins

Ran Rosen, Ayelet Sacher, Nelia Shechter, Dörte Becher, Knut Büttner, Dvora Biran, Michael Hecker, Eliora Z. Ron

Research output: Contribution to journalArticlepeer-review

Abstract

Proteomics based on two-dimensional (2-D) gel electrophoresis of proteins followed by spot identification with mass spectrometry is a commonly used method for physiological studies. Physiological proteomics requires 2-D reference maps, on which most of the main proteins are identified. We present a reference map for the bacterial plant pathogen Agrobacterium tumefaciens proteins, which contains more than 300 entries with an isoelectric point (p/) between 4 and 7. The quantitative study of the proteins in the analytical window of the master gel demonstrated unique features, in comparison with other bacteria. In addition, a theoretical analysis of several protein parameters was performed and compared with the experimental results. A comparison of the theoretical molecular weight (MW) of the proteins and their theoretical p/ with their vertical and horizontal migration distances, respectively, pointed out the existence of several proteins that strongly diverted from the graph trend-line. These proteins were clearly subjected to post-translational modifications, which changed their p/ and/or MW. Additional support for post-translational modifications comes from the identification of multiple spots of the same gene products. Post-translational modifications appear to be more common than expected, at least for soluble proteins, as more than 10% of the proteins were associated with multiple spots.

Original languageEnglish
Pages (from-to)1061-1073
Number of pages13
JournalProteomics
Volume4
Issue number4
DOIs
StatePublished - Apr 2004

Keywords

  • Mass spectrometry
  • Post-translational modifications
  • Rhizobia
  • Stress response

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