Quantitation of yeast cell wall proteins (NWO-CW) in purchasing the nano-HPLC. Frans Klis acknowledges ﬁnancial support from the European Com- mission (STREP FungWall grant, contract LSHB-CT-2004- 511952). Statement Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation. References Aguilar-Uscanga B & Franc¸ois JM (2003) A study of the yeast cell wall composition and structure in response to growth conditions and mode of cultivation. Lett Appl Microbiol 37: 268–274. Baba M & Osumi M (1987) Transmission and scanning electron microscopic examination of intracellular organelles in freeze- substituted Kloeckera and Saccharomyces cerevisiae yeast cells. J Electron Microsc Tech 5: 249–261. Brul S, King A, van der Vaart JM, Chapman J, Klis FM & Verrips CT (1997) The incorporation of mannoproteins in the cell wall of S. cerevisiae and ﬁlamentous Ascomycetes. Antonie Van Leeuwenhoek 72: 229–237. Cabib E, Blanco N, Grau C, Rodrı´quez-Pen˜a JM & Arroyo J (2007) Crh1p and Crh2p are required for the cross-linking of chitin to b(1–6)glucan in the Saccharomyces cerevisiae cell wall. Mol Microbiol 63: 921–935. Caro LH, Tettelin H, Vossen JH, Ram AFJ, van den Ende H & Klis FM (1997) In silicio identiﬁcation of glycosyl- phosphatidylinositol-anchored plasma-membrane and cell wall proteins of Saccharomyces cerevisiae. Yeast 13: 1477–1489. Conzelmann A, Riezman H, Desponds C & Bron C (1988) A major 125-kd membrane glycoprotein of Saccharomyces cerevisiae is attached to the lipid bilayer through an inositol- containing phospholipid. Embo J 7: 2233–2240. Damveld RA, Arentshorst M, VanKuyk PA, Klis FM, van den Hondel CAMJJ & Ram AFJ (2005) Characterisation of CwpA, a putative glycosylphosphatidylinositol-anchored cell wall mannoprotein in the ﬁlamentous fungus Aspergillus niger. Fungal Genet Biol 42: 873–885. De Groot PWJ, Ruiz C, Va´zquez de Aldana CR et al. (2001) A genomic approach for the identiﬁcation and classiﬁcation of genes involved in cell wall formation and its regulation in Saccharomyces cerevisiae. Comp Funct Genomics 2: 124–142. De Groot PWJ, de Boer AD, Cunningham J, Dekker HL, de Jong L, Hellingwerf KJ, de Koster CG & Klis FM (2004) Proteomic analysis of Candida albicans cell walls reveals covalently bound carbohydrate-active enzymes and adhesins. Eukaryot Cell 3: 955–965. De Groot PWJ, Yin QY, Weig M, Sosinska GJ, Klis FM & de Koster CG (2007) Mass spectrometric identiﬁcation of covalently bound cell wall proteins from the ﬁssion yeast Schizosaccharomyces pombe. Yeast 24: 267–278. FEMS Yeast Res 7 (2007) 887–896 895 De Nobel JG, Klis FM, Priem J, Munnik T & van den Ende H (1990) The glucanase-soluble mannoproteins limit cell wall porosity in Saccharomyces cerevisiae. Yeast 6: 491–499. Ecker M, Deutzmann R, Lehle L, Mrsa V & Tanner W (2006) PIR- proteins of Saccharomyces cerevisiae are attached to b-1,3- glucan by a new protein–carbohydrate linkage. J Biol Chem 281: 11523–11529. Fasman GD (1976) CRC Handbook of Biochemistry and Molecular Biology: Proteins, I. CRC Press, Boca Raton, Florida. Ghaemmaghami S, Huh WK, Bower K, Howson RW, Belle A, Dephoure N, O’Shea EK & Weissman JS (2003) Global analysis of protein expression in yeast. Nature 425: 737–741. Hamada K, Terashima H, Arisawa M, Yabuki N & Kitada K (1999) Amino acid residues in the o-minus region participate in cellular localization of yeast glycosylphosphatidylinositol- attached proteins. J Bacteriol 181: 3886–3889. Henrissat B & Davies G (1997) Structural and sequence-based classiﬁcation of glycoside hydrolases. Curr Opin Struct Biol 7: 637–644. Jung US & Levin DE (1999) Genome-wide analysis of gene expression regulated by the yeast cell wall integrity signalling pathway. Mol Microbiol 34: 1049–1057. Kapteyn JC, Ram AFJ, Groos EM, Kollar R, Montijn RC, Van Den Ende H, Llobell A, Cabib E & Klis FM (1997) Altered extent of cross-linking of b1,6-glucosylated mannoproteins to chitin in Saccharomyces cerevisiae mutants with reduced cell wall b1, 3-glucan content. J Bacteriol 179: 6279–6284. Kapteyn JC, ter Riet B, Vink E, Blad S, De Nobel H, Van Den Ende H & Klis FM (2001) Low external pH induces HOG1- dependent changes in the organization of the Saccharomyces cerevisiae cell wall. Mol Microbiol 39: 469–479. Klis FM, Boorsma A & de Groot PWJ (2006) Cell wall construction in Saccharomyces cerevisiae. Yeast 23: 185–202. Lagorce A, Hauser NC, Labourdette D, Rodriguez C, Martin- Yken H, Arroyo J, Hoheisel JD & Francois J (2003) Genome- wide analysis of the response to cell wall mutations in the yeast Saccharomyces cerevisiae. J Biol Chem 278: 20345–20357. Lehninger AL, Nelson DL & Cox MM (1993) Principles of Biochemistry. Worth Publishers, New York. Levin DE (2005) Cell wall integrity signaling in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 69: 262–291. Meijer HJ, van de Vondervoort PJ, Yin QY, de Koster CG, Klis FM, Govers F & de Groot PWJ (2006) Identiﬁcation of cell wall- associated proteins from Phytophthora ramorum. Mol Plant Microbe Interact 19: 1348–1358. Montijn RC, Van Wolven P, De Hoog S & Klis FM (1997) b-Glucosylated proteins in the cell wall of the black yeast Exophiala (Wangiella) dermatitidis. Microbiology 143: 1673–1680. Mouyna I, Fontaine T, Vai M, Monod M, Fonzi WA, Diaquin M, Popolo L, Hartland RP & Latge´ JP (2000) Glycosylphosphatidylinositol-anchored glucanosyltransferases play an active role in the biosynthesis of the fungal cell wall. J Biol Chem 275: 14882–14889. c 2007 The Authors Journal compilation c 2007 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd.