Cell wall protein and glycoprotein constituents of Aspergillus fumigatus that bind to polystyrene may be responsible for the cell surface hydrophobicity of the mycelium.
Cell surface hydrophobicity (CSH) of Aspergillus fumigatus grown both in complex medium (yeast extract/peptone/dextrose; YPD) and minimal (Vogel's N) medium was monitored by assessing attachment of polystyrene microspheres to the cell surface. It was found that mature mycelium was hydrophobic. Treatment of intact mycelium with beta-mercaptoethanol (beta ME) abolished binding of the microspheres to hyphal elements, and coating of the microspheres with beta ME extracts from mycelium inhibited their attachment to intact mycelial cells. A. fumigatus mycelium was tagged in vivo with biotin and treated with beta ME. The beta ME extracts were analysed by SDS-PAGE and Western blotting with both peroxidase-conjugated-ExtrAvidin and concanavalin A (ConA). This procedure allowed identification of cell wall surface proteins and glycoproteins. Rabbit polyclonal antisera were raised against beta ME extracts obtained from cells grown in YPD and Vogel's N media. These antisera defined some major cell-wall-bound antigens. SDS-PAGE and Western blotting analysis of the cell wall material released by beta ME and adsorbed on polystyrene microspheres revealed about 19 protein species with apparent molecular masses ranging from 20 to 70 kDa, and two high-molecular-mass glycoproteins of 115 and 210 kDa. Treatment of cells grown in YPD, but not those grown in Vogel's N medium, with beta ME released a 55 kDa polypeptide able to adsorb to polystyrene microspheres that was detectable with the antisera. The ability to bind to polystyrene particles exhibited by several protein and glycoprotein species released by beta ME treatment suggested that these cell wall moieties possess exposed hydrophobic domains that could be responsible for the CSH of mycelium.
Peñalver
M C
MC
Departamento de Microbiología y Ecología, Facultad de Farmacia, Universitat de València, Spain.
Casanova
M
M
Martínez
J P
JP
Gil
M L
ML
D000818:Animals; D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D001232:Aspergillus fumigatus / Q000254:growth & development / Q000276:immunology / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism; D003470:Culture Media; D005656:Fungal Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008863:Microspheres; D008970:Molecular Weight; D011137:Polystyrenes; D011817:Rabbits; D014867:Water / Q000378:metabolism
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D003470:Culture Media; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011137:Polystyrenes; D014867:Water