Hunting down fungal secretomes using liquid-phase IEF prior to high resolution 2-DE
Delphine Vincent, Marie-Helene Balesdent, Julien Gibon, Stephane Claverol, Delphine Lapaillerie, Anne-Marie Lomenech, Francoise Blaise, Thierry Rouxel, Francis Martin, Marc Bonneu, Joelle Amselem, Victoria Dominguez, Barbara J Howlett, Patrick Wincker, Johann Joets, Marc-Henri Lebrun, Christophe Plomion
ELECTROPHORESIS | WILEY | Published : 2009
The secreted proteins (secretome) of fungi play a key role in interactions of pathogenic and symbiotic fungi with plants. Using the plant pathogenic fungus Leptosphaeria maculans and symbiont Laccaria bicolor grown in culture, we have established a proteomic protocol for extraction, concentration and resolution of the fungal secretome. As no proteomic data were available on mycelium tissues from both L. maculans and L. bicolor, mycelial proteins were studied; they also helped verifying the purity of secretome samples. The quality of protein extracts was initially assessed by both 1-DE and 2-DE using first a broad pH range for IEF, and then narrower acidic and basic pH ranges, prior to 2-DE. ..View full abstract
Awarded by Agence Nationale de la Recherche
The authors are grateful to Matthew D. Wheatley for careful reading of the manuscript, Olivier Langella for helping uploading 2-DE/MS data into PROTICdb, Dr. Franck Salin for his assistance with 1-DE, Dr. Nathalie Ollat for her help with sample lyophilization. This work was funded by the Agence Nationale de la Recherche (ANR-06-BLAN-0399) and Institut National de la Recherche Agronomique (INRA).