Structural analysis of glucoamylase encoded by the STA1 gene of Saccharomyces cerevisiae (var. diastaticus).
The sequence of the STA1-encoded glucoamylase of amylolytic Saccharomyces cerevisiae (var. diastaticus) strains shows two well-defined regions: an amino-terminal part rich in serine and threonine residues and a carboxy-terminal part very similar to the catalytic domain of other fungal glucoamylases. A version of the enzyme in which most of the amino-terminal region was deleted still has glucoamylase activity, indicating that the remaining carboxy-terminal part forms a functional catalytic domain. Homology-based models of the two parts of the protein have been obtained. As expected, the shortened form of the enzyme is very similar to the catalytic domain of related glucoamylases of known structure. However, the amino-terminal part yielded a structure revealing an unexpected similarity to bacterial invasins, suggesting functional connections between several yeast proteins homologous to STA1-encoded glucoamylase and invasins. A characteristic of Saccharomyces glucoamylase in its native form is its extreme degree of glycosylation. Despite its high molecular mass (about 300 kDa), and in contrast with what occurs with other extracellular glycoproteins produced by yeast, the enzyme does not remain attached to the cell wall, being fully and efficiently secreted into the medium, even when it is produced in large amounts by overexpression of its gene.
Adam
Ana Cristina
AC
Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Cientificas, Paterna, Valencia, Spain.
Latorre-García
Lorena
L
Polaina
Julio
J
D000595:Amino Acid Sequence; D020134:Catalytic Domain; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D005087:Glucan 1,4-alpha-Glucosidase / Q000737:chemistry* / Q000235:genetics*; D006031:Glycosylation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*
D004271:DNA, Fungal; D005087:Glucan 1,4-alpha-Glucosidase