Assay for sialidase using erythrocytes and peroxidase-labeled peanut lectin.
A rapid and sensitive sialidase assay method based on peroxidase-labeled peanut lectin (PNA) binding to desialylated erythrocyte is described. Formalinized sheep erythrocytes were used both as a stable substrate for sialidase and as a target for the lectin. In the case of sialidases from Vibrio cholerae and Arthrobacter ureafaciens, a linear relationship was observed between the amount of peroxidase-labeled PNA bound to erythrocytes and the enzyme amount. Binding of the lectin to sialidase-treated erythrocytes was completely prevented in the presence of 25 mM lactose and galactose. The method is particularly useful as a selective assay for sialidase which is active towards gangliosides or sialoglycoproteins, because a mammalian sialidase which is preferentially active towards sialooligosaccharides and sialoglycopeptides is not able to remove sialic acid from erythrocytes.
Nagai
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Yamada
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D000818:Animals; D004912:Erythrocytes / Q000187:drug effects*; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D037102:Lectins; D051379:Mice; D009439:Neuraminidase / Q000494:pharmacology*; D019887:Peanut Agglutinin; D012756:Sheep; D014734:Vibrio cholerae / Q000201:enzymology
D037102:Lectins; D019887:Peanut Agglutinin; D009439:Neuraminidase