Expression of the Cu,Zn superoxide dismutase of Aspergillus fumigatus as determined by immunochemistry and immunoelectron microscopy.
A polyclonal antibody against purified Cu,Zn superoxide dismutase (SOD) from the pathogen Aspergillus fumigatus was raised in a sheep. This antibody recognised purified A. fumigatus SOD, together with a single band of 19 kDa in A. fumigatus cytoplasmic antigen, by immunodevelopment of Western blots. The polyclonal serum did not recognise either the manganese or iron containing forms of the enzyme; however, it was reactive against putative Cu,Zn SODs in other members of the genus Aspergillus. Immunofluorescent staining of A. fumigatus cultures demonstrated expression of the Cu,Zn SOD in conidia and hyphae, with the cell wall staining particularly intensely. Conidiophores were stained in an uniformly intense pattern. Immunoelectron microscopy confirmed that the SOD was present within the hyphal cell wall, although there was also labelling in the cytoplasm. SOD may protect Aspergillus against oxidants produced by immune effector cells and these observations demonstrate that the enzyme is available to perform its antioxidant function within the cell wall.
Hamilton
A J
AJ
Dermatology Unit, St Johns Institute of Dermatology, Guy's Hospital, London, UK.
Holdom
M D
MD
Jeavons
L
L
D000908:Antibodies, Fungal; D000918:Antibody Specificity; D001232:Aspergillus fumigatus / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods*; D016253:Microscopy, Immunoelectron / Q000379:methods; D013482:Superoxide Dismutase / Q000276:immunology / Q000302:isolation & purification*
D000908:Antibodies, Fungal; D013482:Superoxide Dismutase