Abstract
Fungal cell wall is a rigid structure mainly composed of polysaccharides (up to 90 %) and glycoproteins. It is essential for survival of the fungal cells, because it protects them against bursting caused by internal turgor pressure and against mechanical injury. Because of its absence in mammalian cells, it is an attractive target for antifungal agents. Thus, for various reasons, it might be important to know how the cell wall is synthesized, and how to analyze its composition. We provide here information about in vitro analysis of the biosynthetic activities of the main fungal wall and describe some methods for rapid analysis of cell wall composition by using specific enzymatic degradations. We also describe some additional methods that can be occasionally used to analyze fungal wall properties or composition. These methods provide powerful tools to evaluate changes in fungal cell walls and will be useful for screening new compounds for antifungal activity that might cause inhibition of cell wall biosynthesis and/or alter the structure of the fungal cell wall.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Latge JP (2007) The cell wall: a carbohydrate armour for the fungal cell. Mol Microbiol 66:279–290
Cabib E, Bowers B, Sburlati A, Silverman SJ (1988) Fungal cell wall synthesis: the construction of a biological structure. Microbiol Sci 5:370–375
Lesage G, Bussey H (2006) Cell wall assembly in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 70:317–343
Cabib E, Kang MS (1987) Fungal 1,3-b-glucan synthase. Methods Enzymol 138:637–642
Kapteyn JC, Ram AF, Groos EM, Kollar R, Montijn RC, Van Den Ende H et al (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
Kollar R, Reinhold BB, Petráková E, Yeh HJ, Ashwell G, Drgonová J et al (1997) Architecture of the yeast cell wall. b(1-6)-glucan interconnects mannoprotein, b(1-3)-glucan, and chitin. J Biol Chem 272:17762–17775
Kollar R, Petrakova E, Ashwell G, Robbins PW, Cabib E (1995) Architecture of the yeast cell wall. The linkage between chitin and b(1-3)-glucan. J Biol Chem 270:1170–1178
Cabib E, Blanco N, Grau C, Rodriguez-Pena 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
Cabib E, Farkas V, Kosik O, Blanco N, Arroyo J, McPhie P (2008) Assembly of the yeast cell wall. Crh1p and Crh2p act as transglycosylases in vivo and in vitro. J Biol Chem 283:29859–29872
Kapteyn JC, Montijn RC, Vink E, de la Cruz J, Llobell A, Douwes JE, Shimoi H, Lipke PN, Klis FM (1996) Retention of Saccharomyces cerevisiae cell wall proteins through a phosphodiester-linked b-1,3-/b-1,6-glucan heteropolymer. Glycobiology 6:337–345
Klis FM, Boorsma A, De Groot PW (2006) Cell wall construction in Saccharomyces cerevisiae. Yeast 23:185–202
Douglas CM, Marrinan JA, Li W, Kurtz MB (1994) A Saccharomyces cerevisiae mutant with echinocandin-resistant 1,3-b-D-glucan synthase. J Bacteriol 176:5686–5696
Mazur P, Morin N, Baginsky W, El-Sherbeini M, Clemas JA, Nielsen JB et al (1995) Differential expression and function of two homologous subunits of yeast 1,3-b-D-glucan synthase. Mol Cell Biol 15:5671–5681
Pérez P, Ribas JC (2004) Cell wall analysis. Methods 33:245–251
Arellano M, Duran A, Perez P (1996) Rho1 GTPase activates the (1-3)b-D-glucan synthase and is involved in Schizosaccharomyces pombe morphogenesis. EMBO J 15:4584–4591
Qadota H, Python CP, Inoue SB, Arisawa M, Anraku Y, Zheng Y et al (1996) Identification of yeast Rho1p GTPase as a regulatory subunit of 1,3-b-glucan synthase. Science 272:279–281
Katayama S, Hirata D, Arellano M, Pérez P, Toda T (1999) Fission yeast a-glucan synthase Mok1 requires the actin cytoskeleton to localize the sites of growth and plays an essential role in cell morphogenesis downstream of protein kinase C function. J Cell Biol 144:1173–1186
Hoschstenbach F, Klis FM, Van den Ende H, Van Donselaar E, Peters PJ et al (1998) Identification of a putative alpha-glucan synthase essential for cell wall construction and morphogenesis in fission yeast. Proc Natl Acad Sci USA 95:9161–9166
Grun CH, Hochstenbach F, Humbel BM, Verkleij AJ, Sietsma JH, Klis FM et al (2005) The structure of cell wall alpha-glucan from fission yeast. Glycobiology 15:245–257
Vos A, Dekker N, Distel B, Leunissen JA, Hochstenbach F (2007) Role of the synthase domain of Ags1p in cell wall alpha-glucan biosynthesis in fission yeast. J Biol Chem 282:18969–18979
Cabib E, Roh DH, Schmidt M, Crotti LB, Varma A (2001) The yeast cell wall and septum as paradigms of cell growth and morphogenesis. J Biol Chem 276:19679–19682
Roncero C (2002) The genetic complexity of chitin synthesis in fungi. Curr Genet 41:367–378
Lenardon MD, Munro CA, Gow NA (2010) Chitin synthesis and fungal pathogenesis. Curr Opin Microbiol 13:416–423
Bowman SM, Free SJ (2006) The structure and synthesis of the fungal cell wall. Bioessays 28:799–808
Leal JA, Prieto A, Bernabe M, Hawksworth DL (2010) An assessment of fungal wall heteromannans as a phylogenetically informative character in ascomycetes. FEMS Microbiol Rev 34:986–1014
Shematek EM, Braatz JA, Cabib E (1980) Biosynthesis of yeast cell wall. I. Preparation and properties of b(1-3)glucan synthetase. J Biol Chem 255:888–894
Shematek EM, Cabib E (1980) Biosynthesis of yeast cell wall. II. Regulation of b(1-3)glucan synthetase by ATP and GTP. J Biol Chem 255:895–902
Ishiguro J, Saitou A, Durán A, Ribas JC (1997) cps1 +, a Schizosaccharomyces pombe gene homolog of Saccharomyces cerevisiae FKS genes whose mutation confers hypersensitivity to cyclosporin A and papulacandin B. J Bacteriol 179:7653–7662
Martins IM, Cortés JCG, Muñoz J, Moreno MB, Ramos M, Clemente-Ramos JA et al (2011) Differential activities of three families of specific b(1,3)glucan synthase inhibitors in wild-type and resistant strains of fission yeast. J Biol Chem 286:3484–3496
Abe M, Nishida I, Minemura M, Qadota H, Seyama Y, Watanabe T et al (2001) Yeast 1,3-b-glucan synthase activity is inhibited by phytosphingosine localized to the endoplasmic reticulum. J Biol Chem 276:26923–26930
El-Sherbeini M, Clemas JA (1995) Nikkomycin Z supersensitivity of an echinocandin-resistant mutant of Saccharomyces cerevisiae. Antimicrob Agents Chemother 39:200–207
Inoue SB, Takewaki N, Takasuka T, Mio T, Adachi M, Fujii Y et al (1995) Characterization and gene cloning of 1,3-b-D-glucan synthase from Saccharomyces cerevisiae. Eur J Biochem 231:845–854
Kelly R, Register E, Hsu MJ, Kurtz M, Nielsen J (1996) Isolation of a gene involved in 1,3-b-glucan synthesis in Aspergillus nidulans and purification of the corresponding protein. J Bacteriol 178:4381–4391
Mazur P, Baginsky W (1996) In vitro activity of 1,3-b-D-glucan synthase requires the GTP-binding protein Rho1. J Biol Chem 271:14604–14609
Mol PC, Park HM, Mullins JT, Cabib E (1994) A GTP-binding protein regulates the activity of (1,3)-b-glucan synthase, an enzyme directly involved in yeast cell wall morphogenesis. J Biol Chem 269:31267–31274
Thompson JR, Douglas CM, Li W, Jue CK, Pramanik B, Yuan X et al (1999) A glucan synthase FKS1 homolog in Cryptococcus neoformans is single copy and encodes an essential function. J Bacteriol 181:444–453
Wood RL, Miller TK, Wright A, McCarthy P, Taft CS, Pomponi S et al (1998) Characterization and optimization of in vitro assay conditions for (1,3)b-glucan synthase activity from Aspergillus fumigatus and Candida albicans for enzyme inhibition screening. J Antibiot (Tokyo) 51:665–675
Sestak S, Farkas V (2001) In situ assays of fungal enzymes in cells permeabilized by osmotic shock. Anal Biochem 292:34–39
Shedletzky E, Unger C, Delmer DP (1997) A microtiter-based fluorescence assay for (1,3)-b-glucan synthases. Anal Biochem 249:88–93
Ribas JC, Díaz M, Durán A, Pérez P (1991) Isolation and characterization of Schizosaccharomyces pombe mutants defective in cell wall (1-3)b-D-glucan. J Bacteriol 173:3456–3462
El-Sherbeini M, Clemas JA (1995) Cloning and characterization of GNS1: a Saccharomyces cerevisiae gene involved in synthesis of 1,3-b-glucan in vitro. J Bacteriol 177:3227–3234
Kang MS, Cabib E (1986) Regulation of fungal cell wall growth: a guanine nucleotide-binding proteinaceous component required for activity of (1,3)-b-D-glucan synthase. Proc Natl Acad Sci U S A 83:5808–5812
Kondoh O, Tachibana Y, Ohya Y, Arisawa M, Watanabe T (1997) Cloning of the RHO1 gene from Candida albicans and its regulation of b-1,3-glucan synthesis. J Bacteriol 179:7734–7741
Schimoler-O’Rourke R, Renault S, Mo W, Selitrennikoff CP (2003) Neurospora crassa FKS protein binds to the (1,3)b-glucan synthase substrate, UDP-glucose. Curr Microbiol 46:408–412
Crotti LB, Drgon T, Cabib E (2001) Yeast cell permeabilization by osmotic shock allows determination of enzymatic activities in situ. Anal Biochem 292:8–16
Aimanianda V, Clavaud C, Simenel C, Fontaine T, Delepierre M, Latge JP (2009) Cell wall b-(1,6)-glucan of Saccharomyces cerevisiae: structural characterization and in situ synthesis. J Biol Chem 284:13401–13412
Frost DJ, Brandt K, Capobianco J, Goldman R (1994) Characterization of (1,3)-b-glucan synthase in Candida albicans: microsomal assay from the yeast or mycelial morphological forms and a permeabilized whole-cell assay. Microbiology 140:2239–2246
Choi WJ, Cabib E (1994) The use of divalent cations and pH for the determination of specific yeast chitin synthetases. Anal Biochem 219:368–372
Lucero HA, Kuranda MJ, Bulik DA (2002) A nonradioactive, high throughput assay for chitin synthase activity. Anal Biochem 305:97–105
Castro C, Ribas JC, Valdivieso MH, Varona R, del Rey F, Durán A (1995) Papulacandin B resistance in budding and fission yeasts: isolation and characterization of a gene involved in (1,3)b-D-glucan synthesis in Saccharomyces cerevisiae. J Bacteriol 177:5732–5739
Roncero C, Valdivieso MH, Ribas JC, Duran A (1988) Isolation and characterization of Saccharomyces cerevisiae mutants resistant to Calcofluor white. J Bacteriol 170:1950–1954
Algranati ID, Behrens N, Carminatti H, Cabib E (1966) Mannan synthetase from yeast. Methods Enzymol 8:411–416
Gopal PK, Shepherd MG, Sullivan PA (1984) Analysis of wall glucans from yeast, hyphal and germ-tube forming cells of Candida albicans. J Gen Microbiol 130:3295–3301
Gorka-Niec W, Perlinska-Lenart U, Zembek P, Palamarczyk G, Kruszewska JS (2010) Influence of sorbitol on protein production and glycosylation and cell wall formation in Trichoderma reesei. Fungal Biol 114:855–862
Bartnicki-Garcia S (1999) Glucans, walls, and morphogenesis: On the contributions of J. G. H. Wessels to the golden decades of fungal physiology and beyond. Fungal Genet Biol 27:119–127
Perez P, Garcia-Acha I, Duran A (1983) Effect of papulacandin B on the cell wall and growth of Geotrichum lactis. J Gen Microbiol 129:245–250
Sietsma JH, Wessels JG (1977) Chemical analysis of the hyphal wall of Schizophyllum commune. Biochim Biophys Acta 496:225–239
Tomazett PK, Felix CR, Lenzi HL, de Paula Faria F, de Almeida Soares CM, Pereira M (2010) 1,3-b-D-Glucan synthase of Paracoccidioides brasiliensis: recombinant protein, expression and cytolocalization in the yeast and mycelium phases. Fungal Biol 114:809–816
Latge JP (2010) Tasting the fungal cell wall. Cell Microbiol 12:863–872
Manners DJ, Meyer MT (1977) The molecular structures of some glucans from the cell walls of Schizosaccharomyces pombe. Carbohydr Res 57:189–203
Magnelli PE, Cipollo JF, Robbins PW (2005) A glucanase-driven fractionation allows redefinition of Schizosaccharomyces pombe cell wall composition and structure: assignment of diglucan. Anal Biochem 336:202–212
Sugawara T, Takahashi S, Osumi M, Ohno N (2004) Refinement of the structures of cell-wall glucans of Schizosaccharomyces pombe by chemical modification and NMR spectroscopy. Carbohydr Res 339: 2255–2265
Magnelli P, Cipollo JF, Abeijon C (2002) A refined method for the determination of Saccharomyces cerevisiae cell wall composition and b-1,6-glucan fine structure. Anal Biochem 301:136–150
Sugawara T, Sato M, Takagi T, Kamasaki T, Ohno N, Osumi M (2003) In situ localization of cell wall a-1,3-glucan in the fission yeast Schizosaccharomyces pombe. J Electron Microsc (Tokyo) 52:237–242
Bush DA, Horisberger M, Horman I, Wursch P (1974) The wall structure of Schizosaccharomyces pombe. J Gen Microbiol 81:199–206
Cabib E, Durán A (2005) Synthase III-dependent chitin is bound to different acceptors depending on location on the cell wall of budding yeast. J Biol Chem 280:9170–9179
Manners DJ, Masson AJ, Patterson JC (1973) The structure of a b-(1-3)-D-glucan from yeast cell walls. Biochem J 135:19–30
Manners DJ, Masson AJ, Patterson JC, Bjorndal H, Lindberg B (1973) The structure of a b-(1-6)-D-glucan from yeast cell walls. Biochem J 135:31–36
Fontaine T, Simenel C, Dubreucq G, Adam O, Delepierre M, Lemoine J et al (2000) Molecular organization of the alkali-insoluble fraction of Aspergillus fumigatus cell wall. J Biol Chem 275:27594–27607
Montijn RC, van Rinsum J, van Schagen FA, Klis FM (1994) Glucomannoproteins in the cell wall of Saccharomyces cerevisiae contain a novel type of carbohydrate side chain. J Biol Chem 269:19338–19342
Bulawa CE, Slater M, Cabib E, Au-Young J, Sburlati A, Adair WL Jr, Robbins PW (1986) The S. cerevisiae structural gene for chitin synthase is not required for chitin synthesis in vivo. Cell 46:213–225
Reissig JL, Storminger JL, Leloir LF (1955) A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem 217:959–966
Boone C, Sommer SS, Hensel A, Bussey H (1990) Yeast KRE genes provide evidence for a pathway of cell wall b-glucan assembly. J Cell Biol 110:1833–1843
Dijkgraaf GJ, Brown JL, Bussey H (1996) The KNH1 gene of Saccharomyces cerevisiae is a functional homolog of KRE9. Yeast 12:683–692
Dijkgraaf GJ, Abe M, Ohya Y, Bussey H (2002) Mutations in Fks1p affect the cell wall content of b-1,3- and b-1,6-glucan in Saccharomyces cerevisiae. Yeast 19:671–690
Badin J, Jackson C, Schubert M (1953) Improved method for determination of plasma polysaccharides with tryptophan. Proc Soc Exp Biol Med 84:289–291
McKelvy JF, Lee YC (1969) Microheterogeneity of the carbohydrate group of Aspergillus oryzae a-amylase. Arch Biochem Biophys 132:99–110
Lussier M, Sdicu AM, Shahinian S, Bussey H (1998) The Candida albicans KRE9 gene is required for cell wall b-1,6-glucan synthesis and is essential for growth on glucose. Proc Natl Acad Sci U S A 95:9825–9830
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
de Groot PW, Ram AF, Klis FM (2005) Features and functions of covalently linked proteins in fungal cell walls. Fungal Genet Biol 42:657–675
de Groot PW, 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
Klis FM, Brul S, De Groot PW (2010) Covalently linked wall proteins in ascomycetous fungi. Yeast 27:489–493
Mrsa V, Ecker M, Strahl-Bolsinger S, Nimtz M, Lehle L, Tanner W (1999) Deletion of new covalently linked cell wall glycoproteins alters the electrophoretic mobility of phosphorylated wall components of Saccharomyces cerevisiae. J Bacteriol 181:3076–3086
de Groot PW, de Boer AD, Cunningham J, Dekker HL, de Jong L, Hellingwerf KJ et al (2004) Proteomic analysis of Candida albicans cell walls reveals covalently bound carbohydrate-active enzymes and adhesins. Eukaryot Cell 3:955–965
Carotti C, Ragni E, Palomares O, Fontaine T, Tedeschi G, Rodriguez R et al (2004) Characterization of recombinant forms of the yeast Gas1 protein and identification of residues essential for glucanosyltransferase activity and folding. Eur J Biochem 271:3635–3645
Kapteyn JC, ter Riet B, Vink E, Blad S, De Nobel H, Van Den Ende H et al (2001) Low external pH induces HOG1-dependent changes in the organization of the Saccharomyces cerevisiae cell wall. Mol Microbiol 39:469–479
Mrsa V, Seidl T, Gentzsch M, Tanner W (1997) Specific labelling of cell wall proteins by biotinylation. Identification of four covalently linked O-mannosylated proteins of Saccharomyces cerevisiae. Yeast 13:1145–1154
Mrsa V, Tanner W (1999) Role of NaOH-extractable cell wall proteins Ccw5p, Ccw6p, Ccw7p and Ccw8p (members of the Pir protein family) in stability of the Saccharomyces cerevisiae cell wall. Yeast 15:813–820
Ballou L, Hernandez LM, Alvarado E, Ballou CE (1990) Revision of the oligosaccharide structures of yeast carboxypeptidase Y. Proc Natl Acad Sci U S A 87:3368–3372
Calonge TM, Nakano K, Arellano M, Arai R, Katayama S, Toda T et al (2000) Schizosaccharomyces pombe rho2p GTPase regulates cell wall a-glucan biosynthesis through the protein kinase pck2p. Mol Biol Cell 11:4393–4401
Fuglsang CC, Berka RM, Wahleithner JA, Kauppinen S, Shuster JR, Rasmussen G et al (2000) Biochemical analysis of recombinant fungal mutanases. A new family of a1,3-glucanases with novel carbohydrate-binding domains. J Biol Chem 275:2009–2018
Cortés JC, Konomi M, Martins IM, Munoz J, Moreno MB, Osumi M et al (2007) The (1,3)b-D-glucan synthase subunit Bgs1p is responsible for the fission yeast primary septum formation. Mol Microbiol 65:201–217
Bal AM (2010) The echinocandins: three useful choices or three too many? Int J Antimicrob Agents 35:13–18
Chapman SW, Sullivan DC, Cleary JD (2008) In search of the holy grail of antifungal therapy. Trans Am Clin Climatol Assoc 119:197–215; discussion 215–216
Shao PL, Huang LM, Hsueh PR (2007) Recent advances and challenges in the treatment of invasive fungal infections. Int J Antimicrob Agents 30:487–495
Vicente MF, Basilio A, Cabello A, Pelaez F (2003) Microbial natural products as a source of antifungals. Clin Microbiol Infect 9:15–32
Varona R, Pérez P, Durán A (1983) Effect of papulacandin B on b-glucan synthesis in Schizosaccharomyces pombe. FEMS Microbiol Lett 20:243–247
Kitamura A, Higuchi S, Hata M, Kawakami K, Yoshida K, Namba K et al (2009) Effect of b-1,6-glucan inhibitors on the invasion process of Candida albicans: potential mechanism of their in vivo efficacy. Antimicrob Agents Chemother 53:3963–3971
Kitamura A, Someya K, Hata M, Nakajima R, Takemura M (2009) Discovery of a small-molecule inhibitor of b-1,6-glucan synthesis. Antimicrob Agents Chemother 53:670–677
Biely P, Kovarik J, Bauer S (1973) Cell wall formation in yeast. An electron microscopic autoradiographic study. Arch Microbiol 94:356–371
Feldmesser M, Kress Y, Mednick A, Casadevall A (2000) The effect of the echinocandin analogue caspofungin on cell wall glucan synthesis by Cryptococcus neoformans. J Infect Dis 182:1791–1795
Johnson BF, Yoo BY, Calleja GB (1973) Cell division in yeasts: movement of organelles associated with cell plate growth of Schizosaccharomyces pombe. J Bacteriol 115:358–366
Osumi M, Sato M, Ishijima SA, Konomi M, Takagi T, Yaguchi H (1998) Dynamics of cell wall formation in fission yeast, Schizosaccharomyces pombe. Fungal Genet Biol 24:178–206
Roh DH, Bowers B, Schmidt M, Cabib E (2002) The septation apparatus, an autonomous system in budding yeast. Mol Biol Cell 13:2747–2759
Schmidt M, Bowers B, Varma A, Roh DH, Cabib E (2002) In budding yeast, contraction of the actomyosin ring and formation of the primary septum at cytokinesis depend on each other. J Cell Sci 115:293–302
Coluccio A, Bogengruber E, Conrad MN, Dresser ME, Briza P, Neiman AM (2004) Morphogenetic pathway of spore wall assembly in Saccharomyces cerevisiae. Eukaryot Cell 3:1464–1475
Bowers B, Levin G, Cabib E (1974) Effect of polyoxin D on chitin synthesis and septum formation in Saccharomyces cerevisiae. J Bacteriol 119:564–575
Cabib E, Sburlati A, Bowers B, Silverman SJ (1989) Chitin synthase 1, an auxiliary enzyme for chitin synthesis in Saccharomyces cerevisiae. J Cell Biol 108:1665–1672
Shaw JA, Mol PC, Bowers B, Silverman SJ, Valdivieso MH, Durán A et al (1991) The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle. J Cell Biol 114:111–123
Konomi M, Fujimoto K, Toda T, Osumi M (2003) Characterization and behaviour of a-glucan synthase in Schizosaccharomyces pombe as revealed by electron microscopy. Yeast 20:427–438
Osumi M, Yamada N, Kobori H, Taki A, Naito N, Baba M et al (1989) Cell wall formation in regenerating protoplasts of Schizosaccharomyces pombe: study by high resolution, low voltage scanning electron microscopy. J Electron Microsc (Tokyo) 38:457–468
Konomi M, Ishiguro J, Osumi M (2000) Abnormal formation of the glucan network from regenerating protoplasts in Schizosaccharomyces pombe cps8 actin point mutant. J Electron Microsc (Tokyo) 49:569–578
Osumi M (1998) The ultrastructure of yeast: cell wall structure and formation. Micron 29:207–233
Kopecka M, Fleet GH, Phaff HJ (1995) Ultrastructure of the cell wall of Schizosaccharomyces pombe following treatment with various glucanases. J Struct Biol 114:140–152
Sipiczki M, Yamaguchi M, Grallert A, Takeo K, Zilahi E, Bozsik A et al (2000) Role of cell shape in determination of the division plane in Schizosaccharomyces pombe: random orientation of septa in spherical cells. J Bacteriol 182:1693–1701
Osumi M, Yamada N, Yaguchi H, Kobori H, Nagatani T, Sato M (1995) Ultrahigh-resolution low-voltage SEM reveals ultrastructure of the glucan network formation from fission yeast protoplast. J Electron Microsc (Tokyo) 44:198–206
Kobori H, Yamada N, Taki A, Osumi M (1989) Actin is associated with the formation of the cell wall in reverting protoplasts of the fission yeast Schizosaccharomyces pombe. J Cell Sci 94:635–646
Bernard M, Latge JP (2001) Aspergillus fumigatus cell wall: composition and biosynthesis. Med Mycol 39:9–17
Dufrene YF (2010) Atomic force microscopy of fungal cell walls: an update. Yeast 27:465–471
Zhao L, Schaefer D, Xu H, Modi SJ, LaCourse WR, Marten MR (2005) Elastic properties of the cell wall of Aspergillus nidulans studied with atomic force microscopy. Biotechnol Prog 21:292–299
Osumi M, Konomi M, Sugawara T, Takagi T, Baba M (2006) High-pressure freezing is a powerful tool for visualization of Schizosaccharomyces pombe cells: ultra-low temperature and low-voltage scanning electron microscopy and immunoelectron microscopy. J Electron Microsc (Tokyo) 55:75–88
Humbel BM, Konomi M, Takagi T, Kamasawa N, Ishijima SA, Osumi M (2001) In situ localization of b-glucans in the cell wall of Schizosaccharomyces pombe. Yeast 18:433–444
Meikle PJ, Bonig I, Hoogenraad NJ, Clarke AE, Stone BA (1991) The location of (1-3)-b-glucans in the walls of pollen tubes of Nicotiana alata using a (1-3)-b-glucan-specific monoclonal antibody. Planta 185:1–8
Meikle PJ, Hoogenraad NJ, Bonig I, Clarke AE, Stone BA (1994) A (1->3,1->4)-beta-glucan-specific monoclonal antibody and its use in the quantitation and immunocytochemical location of (1->3,1->4)-beta-glucans. Plant J 5:1–9
Montijn RC, Vink E, Muller WH, Verkleij AJ, Van Den Ende H, Henrissat B et al (1999) Localization of synthesis of b1,6-glucan in Saccharomyces cerevisiae. J Bacteriol 181:7414–7420
Adachi Y, Ohno N, Yadomae T (1994) Preparation and antigen specificity of an anti-(1-3)-b-D-glucan antibody. Biol Pharm Bull 17:1508–1512
Reese AJ, Yoneda A, Breger JA, Beauvais A, Liu H, Griffith CL et al (2007) Loss of cell wall a(1-3) glucan affects Cryptococcus neoformans from ultrastructure to virulence. Mol Microbiol 63:1385–1398
Rappleye CA, Eissenberg LG, Goldman WE (2007) Histoplasma capsulatum a-(1,3)-glucan blocks innate immune recognition by the b-glucan receptor. Proc Natl Acad Sci U S A 104:1366–1370
Eissenberg LG, Moser SA, Goldman WE (1997) Alterations to the cell wall of Histoplasma capsulatum yeasts during infection of macrophages or epithelial cells. J Infect Dis 175:1538–1544
Kugler S, Schurtz Sebghati T, Groppe Eissenberg L, Goldman WE (2000) Phenotypic variation and intracellular parasitism by Histoplasma capsulatum. Proc Natl Acad Sci U S A 97:8794–8798
Gonzalez M, Goddard N, Hicks C, Ovalle R, Rauceo JM, Jue CK et al (2010) A screen for deficiencies in GPI-anchorage of wall glycoproteins in yeast. Yeast 27:583–596
Cabib E, Bowers B, Roberts RL (1983) Vectorial synthesis of a polysaccharide by isolated plasma membranes. Proc Natl Acad Sci U S A 80:3318–3321
Molano J, Bowers B, Cabib E (1980) Distribution of chitin in the yeast cell wall. An ultrastructural and chemical study. J Cell Biol 85:199–212
Roberts RL, Bowers B, Slater ML, Cabib E (1983) Chitin synthesis and localization in cell division cycle mutants of Saccharomyces cerevisiae. Mol Cell Biol 3:922–930
Acknowledgements
We thank D. Posner for language revision. This work was supported by grants BFU2010-15641 and BIO2009-10597 from the Dirección General de Investigación, MICINN, Spain, and grant CSI038A11-2 from the Junta de Castilla y León, Spain.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this chapter
Cite this chapter
Pérez, P., Ribas, J.C. (2013). Fungal Cell Wall Analysis. In: Gupta, V., Tuohy, M., Ayyachamy, M., Turner, K., O’Donovan, A. (eds) Laboratory Protocols in Fungal Biology. Fungal Biology. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-2356-0_12
Download citation
DOI: https://doi.org/10.1007/978-1-4614-2356-0_12
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4614-2355-3
Online ISBN: 978-1-4614-2356-0
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)