Open Access

Comparative proteomics using 2-D gel electrophoresis and mass spectrometry as tools to dissect stimulons and regulons in bacteria with sequenced or partially sequenced genomes

  • Sergio Encarnación1Email author,
  • Magdalena Hernández1,
  • Gabriel Martínez-Batallar1,
  • Sandra Contreras1,
  • María del Carmen Vargas1 and
  • Jaime Mora1
Biological Procedures Online7:710117

https://doi.org/10.1251/bpo110

Received: 26 July 2004

Accepted: 18 July 2005

Abstract

We propose two-dimensional gel electrophoresis (2-DE) and mass spectrometry to define the protein components of regulons and stimulons in bacteria, including those organisms where genome sequencing is still in progress. The basic 2-DE protocol allows high resolution and reproducibility and enables the direct comparison of hundreds or even thousands of proteins simultaneously. To identify proteins that comprise stimulons and regulons, peptide mass fingerprint (PMF) with matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS) analysis is the first option and, if results from this tool are insufficient, complementary data obtained with electrospray ionization tandem-MS (ESI-MS/MS) may permit successful protein identification. ESI-MS/MS and MALDI-TOF-MS provide complementary data sets, and so a more comprehensive coverage of a proteome can be obtained using both techniques with the same sample, especially when few sequenced proteins of a particular organism exist or genome sequencing is still in progress.

Indexing terms

Spectrum Analysis, Mass Rhizobium Electrophoresis, Gel, Two-Dimensional

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