Determination of the Sialic Acid Linkage Specificity of Sialidases Using Lectins in a Solid Phase Assay

✍ Scribed by F. Rogerieux; M. Belaise; H. Terzidistrabelsi; A. Greffard; Y. Pilatte; C.R. Lambre

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 431 KB
• Volume: 211
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1993.1257

✦ Synopsis

A procedure for the determination of activity and linkage specificity of sialidases is deseribed. The sialoglycoprotein fetuin is coated onto a microtiter plate and incubated with sialidases from different sources. Enzymatic activities and linkage specificities are then determined by a sandwich method which measured the binding of different lectins to fetuin. The lectins used were peanut agglutinin (PNA) from Arachis hypogaea, which binds specifically the galactose (\beta-1-3-N)-acetylgalactosamine structures that are unmasked following sialidase treatment of fetuin, the lectins from Sambucus nigra (SNA) and Maackia amurensis (MAA) that are specific for (\alpha-2-6) and (\alpha-2-3) bound sialic acids, respectively, and the slug agglutinin from Limax flavus (LFA) that is specific for (\boldsymbol{N})-acetyl and (\boldsymbol{N})-glycolyl neuraminic acids. Increased PNA and decreased LFA, SNA, and MAA lectin binding correlated with sialidase-induced desialylation of the substrate. In this report, the assay was used to determine the activities and specificities of influenza, Vibrio cholerae, and Arthrobacter ureafaciens sialidases. 1993 Academic Press, Inc.