A method was developed for isolating and sequencing proteins present in the extracellular matrix (ECM) of germlings and hyphae of
filamentous fungi. Surface proteins of the cereal pathogen Bipolaris sorokiniana were labelled with a membrane impermeable biotinylating
agent and extracted using a glycine^HCl buffer. Extracted proteins were purified by affinity binding to streptavidin-conjugated magnetic
beads or by two-dimensional gel electrophoresis. Four of the biotinylated proteins from the ECM of B. sorokiniana were isolated, in gel
digested with trypsin and partly sequenced by tandem mass spectrometry. No significant sequence similarities to proteins in databases were
obtained. ß 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.