Open AccessAnalysis of proteins in the extracellular matrix of the plant pathogenic fungus Bipolaris sorokiniana using 2‐D gel electrophoresis and MS/MS
Author(s) -
Apoga Dace,
Ek Bo,
Tunlid Anders
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10596.x
Subject(s) - bipolaris , gel electrophoresis , fungus , extracellular matrix , two dimensional gel electrophoresis , electrophoresis , biology , botany , microbiology and biotechnology , proteomics , chemistry , chromatography , biochemistry , gene
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.