Analysis of proteins in the extracellular matrix of the plant pathogenic fungus Bipolaris sorokiniana using 2-D gel electrophoresis and MS/MS

A method was developed for isolating and sequencing proteins present in the extracellular matrix (ECM) of germlings and hyphae of filamentous fungi, S urface proteins of the cereal pathogen Bipolaris sorokiniana were labelled with a membrane impermeable biotinylating agent and extracted using a glyci ne-HCl buffer. Extracted proteins were purified by affinity binding to stre ptavidin-conjugated magnetic beads or by two-dimensional gel electrophoresi s. Four of the biotinylated proteins from the ECM of B. sorokiniana were is olated, in gel digested with trypsin and partly sequenced by tandem mass sp ectrometry. No significant sequence similarities to proteins in databases w ere obtained. (C) 2001 Federation of European Microbiological Societies. Pu blished by Elsevier Science B.V. All rights, reserved.