title: Elemental biochemical analysis of the polysaccharides in the extracellular matrix of the yeast Saccharomyces cerevisiae.
abstract: In yeast multicellular aggregates, such as biofilms and colonies, cells are supported by a yeast extracellular matrix (yECM) of glycosidic nature, the composition of which is mostly unknown. Saccharomyces cerevisiae ECM was produced, extracted and partitioned. An analytical-grade pure glycoside fraction was obtained, fractionated by anionic exchange liquid chromatography and analyzed by gas chromatography-mass spectrometry and polyacrylamide gel electrophoresis. Two different molecular weight polysaccharides were found, composed of glucose, mannose and small relative amounts of galactose. One of the polysaccharides had a low molecular weight, compatible with the association with glycoproteins abundantly occurring in yECM. In addition, these polysaccharide species were separated by diaminopropane agarose gel electrophoresis and induced metachromatic shift, suggesting chemical substitution, which was corroborated by anticoagulation activity. This was shown to be associated with the double deletion of the yeast homologues of the mammalian Hedgehog modulators Hhatl and Hhat, respectively yeast Gup1 and Gup2. These results pioneer the study of the molecular biology of the ECM supporting S. cerevisiae multicellular aggregates such as biofilms.
authors.affiliation: Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Portugal.
authors.forename: Fábio
authors.initials: F
authors.lastname: Faria-Oliveira
authors.affiliation: Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Carvalho
authors.affiliation: Laboratory of Glycoconjugates Biochemistry and Cellular Biology, Federal University of Rio de Janeiro, Campus of Macaé, RJ, Brazil.|Laboratory of Glycoconjugates Biochemistry and Cellular Biology, Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil.
authors.forename: Celso L R
authors.initials: CL
authors.lastname: Belmiro
authors.affiliation: Laboratory of Glycoconjugates Biochemistry and Cellular Biology, Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Ramalho
authors.affiliation: Laboratory of Glycoconjugates Biochemistry and Cellular Biology, Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil.
authors.forename: Mauro
authors.initials: M
authors.lastname: Pavão
authors.affiliation: Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Portugal.
authors.forename: Cândida
authors.initials: C
authors.lastname: Lucas
authors.affiliation: Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Portugal.
authors.forename: Célia
authors.initials: C
authors.lastname: Ferreira
mesh_terms: D005109:Extracellular Matrix / Q000737:chemistry*; D062610:Fungal Polysaccharides / Q000032:analysis / Q000737:chemistry*; D005690:Galactose / Q000032:analysis; D005947:Glucose / Q000032:analysis; D008358:Mannose / Q000032:analysis; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000737:chemistry*
chemical_list: D062610:Fungal Polysaccharides; D005947:Glucose; D008358:Mannose; D005690:Galactose
keywords: Extracellular matrix; Polysaccharides; Yeast