title: Proteome analysis of Aspergillus fumigatus identifies glycosylphosphatidylinositol-anchored proteins associated to the cell wall biosynthesis.
abstract: Previous studies in Aspergillus fumigatus (Mouyna I., Fontaine T., Vai M., Monod M., Fonzi W. A., Diaquin M., Popolo L., Hartland R. P., Latgé J.-P, J. Biol. Chem. 2000, 275, 14882-14889) have shown that a glucanosyltransferase playing an important role in fungal cell wall biosynthesis is glycosylphosphatidylinositol (GPI) anchored to the membrane. To identify other GPI-anchored proteins putatively involved in cell wall biogenesis, a proteomic analysis has been undertaken in A. fumigatus and the protein data were matched with the yeast genomic data. GPI-anchored proteins of A. fumigatus were released from membrane preparation by an endogenous GPI-phospholipase C, purified by liquid chromatography and separated by two-dimensional electrophoresis. They were characterized by their peptide mass fingerprint through matrix-assisted laser desorption/ionization-time of flight-(MALDI-TOF)-mass spectrometry and by internal amino acid sequencing. Nine GPI-anchored proteins were identified in A. fumigatus. Five of them were homologs of putatively GPI-anchored yeast proteins (Csa1p, Crh1p, Crh2p, Ecm33p, Gas1p) of unknown function but shown by gene disruption analysis to play a role in cell wall morphogenesis. In addition, a comparative study performed with chitin synthase and glucanosyl transferase mutants of A. fumigatus showed that a modification of the growth phenotype seen in these mutants was associated to an alteration of the pattern of GPI-anchored proteins. These results suggest that GPI-anchored proteins identified in this study are involved in A. fumigatus cell wall organization.
authors.affiliation: Biochemistry Department, Aventis-Hoechst Marion Roussel, France. jean-michel.bruneau@aventis.com
authors.forename: J M
authors.initials: JM
authors.lastname: Bruneau
authors.forename: T
authors.initials: T
authors.lastname: Magnin
authors.forename: E
authors.initials: E
authors.lastname: Tagat
authors.forename: R
authors.initials: R
authors.lastname: Legrand
authors.forename: M
authors.initials: M
authors.lastname: Bernard
authors.forename: M
authors.initials: M
authors.lastname: Diaquin
authors.forename: C
authors.initials: C
authors.lastname: Fudali
authors.forename: J P
authors.initials: JP
authors.lastname: Latgé
mesh_terms: D000818:Animals; D001232:Aspergillus fumigatus / Q000737:chemistry* / Q000235:genetics; D002473:Cell Wall; D002853:Chromatography, Liquid / Q000379:methods; D003429:Cross Reactions; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005656:Fungal Proteins / Q000032:analysis*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis*; D009154:Mutation; D010449:Peptide Mapping / Q000379:methods; D020543:Proteome / Q000032:analysis*; D011817:Rabbits; D012685:Sepharose; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D005656:Fungal Proteins; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D020543:Proteome; D012685:Sepharose