Identification of the catalytic residues of the first family of beta(1-3)glucanosyltransferases identified in fungi.
A new family of glycosylphosphatidylinositol-anchored beta(1-3)glucanosyltransferases (Gelp), recently identified and characterized in the filamentous fungus Aspergillus fumigatus, showed functional similarity to the Gas/Phr/Epd protein families, which are involved in yeast morphogenesis. Sequence comparisons and hydrophobic cluster analysis (HCA) showed that all the Gas/Phr/Epd/Gel proteins belong to a new family of glycosylhydrolases, family 72. We confirmed by site-directed mutagenesis and biochemical analysis that the two conserved glutamate residues (the putative catalytic residues of this family, as determined by HCA) are involved in the active site of this family of glycosylhydrolases.
Mouyna
I
I
Institut Pasteur, Laboratoire des Aspergillus, 25 rue du Docteur Roux, 75724 Paris Cédex 15, France. imouyna@pasteur.fr
Monod
M
M
Fontaine
T
T
Henrissat
B
B
Léchenne
B
B
Latgé
J P
JP
D000595:Amino Acid Sequence; D001232:Aspergillus fumigatus / Q000201:enzymology*; D001665:Binding Sites; D015153:Blotting, Western; D020134:Catalytic Domain; D017124:Conserved Sequence / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D018698:Glutamic Acid / Q000235:genetics / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D009154:Mutation / Q000235:genetics; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D005656:Fungal Proteins; D017261:Glycosylphosphatidylinositols; D011994:Recombinant Proteins; D018698:Glutamic Acid; C117939:1,3-beta-glucanosyltransferase; D004693:Glucan Endo-1,3-beta-D-Glucosidase