Current Genetics

, Volume 49, Issue 3, pp 178–189

Optimisation of a 2-D gel electrophoresis protocol for the human-pathogenic fungus Aspergillus fumigatus

  • Olaf Kniemeyer
  • Franziska Lessing
  • Olaf Scheibner
  • Christian Hertweck
  • Axel A. Brakhage
Research Article

DOI: 10.1007/s00294-005-0047-9

Cite this article as:
Kniemeyer, O., Lessing, F., Scheibner, O. et al. Curr Genet (2006) 49: 178. doi:10.1007/s00294-005-0047-9

Abstract

Aspergillus fumigatus is the most important airborne fungal pathogen causing life-threatening infections in immunosuppressed patients. One of the important questions concerning A. fumigatus is the identification of pathogenicity determinants. To obtain a comprehensive overview about the proteins produced at different physiological conditions that are related to the infectious process a proteomic approach has been applied. Here, 2-D gel electrophoresis for filamentous fungi was optimised concerning removal of interfering compounds, protein extraction and separation methods. A trichloroacetic acid-based precipitation method of proteins with their subsequent solubilisation by the use of a combination of CHAPS with a second sulfobetaine detergent gave the best results. The optimised protocol was evaluated by the analysis of the proteomes of A. fumigatus grown on two different carbon sources, i.e., glucose and ethanol. Carbon catabolite repression has not been studied in detail at the protein level in A. fumigatus yet. In addition, growth on ethanol leads to activation of the glyoxylate cycle which was shown to be essential for pathogenesis in bacteria and fungi. In A. fumigatus, differential patterns of enzymes of the gluconeogenesis, glyoxylate cycle and ethanol degradation pathway during growth on glucose and ethanol were observed.

Keywords

Aspergillus fumigatusProteome analysis2-D gel electrophoresisCarbon catabolite repressionGlyoxylate cycle

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Olaf Kniemeyer
    • 1
    • 2
  • Franziska Lessing
    • 1
    • 2
  • Olaf Scheibner
    • 3
  • Christian Hertweck
    • 3
  • Axel A. Brakhage
    • 1
    • 2
  1. 1.Department of Microbiology and Molecular BiologyFriedrich-Schiller-UniversityJenaGermany
  2. 2.Department of Molecular and Applied MicrobiologyLeibniz Institute for Natural Product Research and Infection Biology (HKI)JenaGermany
  3. 3.Department of Biostructure ChemistryLeibniz Institute for Natural Product Research and Infection Biology (HKI)JenaGermany