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Carbohydrate profiling of fungal cell wall surface glycoconjugates of Trichophyton tonsurans and other keratinophilic filamentous fungi using lectins
Author(s) -
Leal André Ferraz Goiana,
de Lima Neto Reginaldo Gonçalves,
Macêdo Danielle Patrícia Cerqueira,
Beltrão Eduardo Isidoro Carneiro,
Neves Rejane Pereira
Publication year - 2011
Publication title -
mycoses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.13
H-Index - 69
eISSN - 1439-0507
pISSN - 0933-7407
DOI - 10.1111/j.1439-0507.2011.02026.x
Subject(s) - glycoconjugate , lectin , concanavalin a , wheat germ agglutinin , microbiology and biotechnology , agglutinin , biochemistry , mannose , biology , cell wall , chemistry , in vitro
Summary Various researchers have concluded that lectins are useful reagents for the study of fungal cell wall surface glycoconjugates. In this study, we evaluated the expression of N ‐acetyl‐ d ‐glucosamine, l ‐fucose, d ‐galactose and glucose/mannose on the cell wall surface of Trichophyton tonsurans and other keratinophilic filamentous fungi, using a simple lectin‐binding protocol. The fungal cultures used were isolated from soils obtained from public parks by the hair‐bait technique. The lectin assays used concanavalin A (Con A), wheat germ agglutinin (WGA), Ulex europeus agglutinin I (UEA‐I) and peanut agglutinin (PNA), all conjugated with horseradish peroxidase. Adhesive tape was placed sticky‐side down over the fungal colony, gently pressed and then removed. The fungal‐tape samples were incubated with the lectin for 1 h at 4 °C. Lectin binding was visualised using 3,3‐diaminobendizine (DAB) and hydrogen peroxidase. There was a high expression of N ‐acetyl‐ d ‐glucosamine on the cell wall surface of all fungi species tested, whereas the expression of l ‐fucose, d ‐galactose and glucose/mannose demonstrated inter‐specific variations. The lectin‐binding assay presented in this article eliminates many of the laborious steps involved in other protocols. The amount and quality of the mycelium and spores immobilised by the adhesive tapes were suitable for obtaining the carbohydrate profile in glycoconjugates of the cell wall surface of filamentous fungi.