Lectin blot studies on proteins of Myxobolus cerebralis, the causative agent of whirling disease.
It is known that Myxobolus cerebralis antigens, both surficial and secreted, are key modulators for, or targets of, host immune system compounds. We undertook SDS-PAGE glycoprotein characterisation of M. cerebralis developmental stages isolated from infected rainbow trout and Western blot analyses using selected biotin-labelled plant lectins (GSA-I, PHA-E, SJA, GSA-II) and anti-triactinomyxon polyclonal antibodies. Glycoproteins were isolated with lectin-affinity chromatography, and prominent bands were characterised by matrix-assisted laser desorption/ionisation-mass spectrometry (MALDI/MS). We identified glycoproteins of M. cerebralis myxospores that contained carbohydrate motifs reactive with Phaseolus vulgaris erythroagglutinin (proteins 20 to 209 kDa, PHA-E), Sophora japonica agglutinin (proteins 7 to 70 kDa, SJA), Griffonia simplicifolia Agglutinin I (proteins 10 to 209 kDa, GSA-I) and G. simplicifolia Agglutinin II (proteins 5 to 40 kDa, GSA-II). Mcgp33, a glycoprotein isolated by lectin-affinity chromatography, was reactive with SJA (about 33 kDa). Antiserum produced against M. cerebralis triactinomyxons was found to have differences in the antigenicity of isolated glycoproteins from both M. cerebralis myxospores and actinospores. We also demonstrated modified antigen expression, especially involving the glycoprotein Mcgp33, in different developmental stages of M. cerebralis.
Knaus
Martin
M
Institute for Zoology, Fish Biology and Fish Diseases, Faculty of Veterinary Science, University of Munich, Kaulbachstrasse 39, 80539 Munich, Germany.
El-Matbouli
Mansour
M
D000818:Animals; D000953:Antigens, Protozoan / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D056890:Eukaryota / Q000254:growth & development / Q000378:metabolism*; D006023:Glycoproteins / Q000302:isolation & purification* / Q000378:metabolism; D015151:Immunoblotting; D013058:Mass Spectrometry; D017686:Oncorhynchus mykiss / Q000469:parasitology*; D037121:Plant Lectins
D000953:Antigens, Protozoan; D006023:Glycoproteins; D037121:Plant Lectins