Anionogenic groups and surface sialoglycoconjugate structures of yeast forms of the human pathogen Paracoccidioides brasiliensis

The surface anionogenic groups and sialoglycoconjugate structures of Paracoccidioides brasiliensis yeast forms were analysed by cell microelectrophoresis, binding assays with lectins and viral particles, ultrastructural cytochemistry, enzymic digestion and flow cytofluorimetry. P. brasiliensis yeast forms, particularly the budding primordia, reacted strongly with cationized ferritin. Binding assays showed that the reaction with sialic-acid-specific Limax flavus lectin (LFA) was distributed over the entire P. brasiliensis cell wall. Treatment of yeast forms with neuraminidase significantly reduced their negative surface charge and LFA labelling, which suggests that sialic acid residues are major anionogenic groups exposed on the P. brasiliensis surface. Furthermore, after neuraminidase treatment, labelling with Arachis hypogaea (peanut) agglutinin increased due to unmasking of subterminal beta-D-galactopyranosyl residues. The sialic acid linkages to galactose are alpha 2,6 and alpha 2,3 as assessed, respectively, by fungal attachment to M1/5 and M1/5 HS8 strains of influenza A virus and binding of Sambucus niger and Maackia amurensis agglutinins. The alpha 2,6 linkage clearly predominated in both experiments. Flow cytofluorimetry analysis revealed the heterogenicity of P. brasiliensis yeast cell populations, which comprised young and mature budding yeasts. Both express binding sites to LFA and Limulus polyphemus agglutinin.UFRJ, Inst Microbiol Prof Paulo de Goes, Ilha Fundao, BR-21941590 Rio De Janeiro, BrazilUFRJ, Inst Biofis Carlos Chagas Filho, Ilha Fundao, BR-21949970 Rio De Janeiro, BrazilUniv Estadual Norte Fluminense, Ctr Biociecias & Biotecnol, BR-28015620 Campos, RJ, BrazilUniv Fed Sao Paulo, Disciplina Biol Celular, BR-04023062 Sao Paulo, BrazilUniv Fed Sao Paulo, Disciplina Biol Celular, BR-04023062 Sao Paulo, BrazilWeb of Scienc