Mass spectrometric identification of covalently bound cell wall proteins from the fission yeast Schizosaccharomyces pombe.
The cell wall of Schizosaccharomyces pombe is bilayered, consisting of an inner layer of mainly polysaccharides and an outer layer of galactomannoproteins. We present a detailed analysis of the cell wall proteome. Six covalently-bound cell wall proteins (CWPs) were identified using tandem mass spectrometry, including four predicted GPI-dependent CWPs (Gas1p, Gas5p, Ecm33p and Pwp1p) and two alkali-sensitive CWPs (Psu1p and Asl1p). Gas1p and Gas5p belong to glycoside hydrolase family 72, and are believed to be involved in 1,3-beta-glucan elongation. Ecm33p belongs to a ubiquitous fungal protein family with an unknown but crucial function in cell wall integrity. Pwp1p is an abundant protein with an unknown but probably non-enzymatic function. All four CWPs were present in HF-pyridine extracts, indicating that they are linked via a phosphodiester bridge to the glucan network. Psu1p is a homologue of the Saccharomyces cerevisiae Sun family, whereas Asl1p has no homologues in S. cerevisiae but is related to Aspergillus fumigatus and Ustilago maydis proteins. Finally, although the protein content of Sz. pombe cell walls is only slightly less than in S. cerevisiae and Candida albicans, the amount of carbohydrate added to the proteins was found to be two- to three-fold decreased, consistent with earlier reported differences in outer chain N-glycosylation.
de Groot
Piet W J
PW
SILS-Biomolecular Mass Spectrometry, University of Amsterdam, The Netherlands. P.W.J.deGroot@uva.nl
Yin
Qing Yuan
QY
Weig
Michael
M
Sosinska
Grazyna J
GJ
Klis
Frans M
FM
de Koster
Chris G
CG
D000468:Alkalies / Q000494:pharmacology; D000595:Amino Acid Sequence; D002473:Cell Wall / Q000737:chemistry* / Q000378:metabolism; D019295:Computational Biology; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D020543:Proteome; D012568:Schizosaccharomyces / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000032:analysis*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
D000468:Alkalies; D017261:Glycosylphosphatidylinositols; D010455:Peptides; D020543:Proteome; D029702:Schizosaccharomyces pombe Proteins; D012967:Sodium Dodecyl Sulfate; D014357:Trypsin