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. 2000 May 1;347(Pt 3):741–747.

Identification of the catalytic residues of the first family of beta(1-3)glucanosyltransferases identified in fungi.

I Mouyna 1, M Monod 1, T Fontaine 1, B Henrissat 1, B Léchenne 1, J P Latgé 1
PMCID: PMC1221011  PMID: 10769178

Abstract

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.

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Selected References

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  1. Borg-von Zepelin M., Beggah S., Boggian K., Sanglard D., Monod M. The expression of the secreted aspartyl proteinases Sap4 to Sap6 from Candida albicans in murine macrophages. Mol Microbiol. 1998 May;28(3):543–554. doi: 10.1046/j.1365-2958.1998.00815.x. [DOI] [PubMed] [Google Scholar]
  2. Callebaut I., Labesse G., Durand P., Poupon A., Canard L., Chomilier J., Henrissat B., Mornon J. P. Deciphering protein sequence information through hydrophobic cluster analysis (HCA): current status and perspectives. Cell Mol Life Sci. 1997 Aug;53(8):621–645. doi: 10.1007/s000180050082. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ducros V., Czjzek M., Belaich A., Gaudin C., Fierobe H. P., Belaich J. P., Davies G. J., Haser R. Crystal structure of the catalytic domain of a bacterial cellulase belonging to family 5. Structure. 1995 Sep 15;3(9):939–949. doi: 10.1016/S0969-2126(01)00228-3. [DOI] [PubMed] [Google Scholar]
  4. Gaboriaud C., Bissery V., Benchetrit T., Mornon J. P. Hydrophobic cluster analysis: an efficient new way to compare and analyse amino acid sequences. FEBS Lett. 1987 Nov 16;224(1):149–155. doi: 10.1016/0014-5793(87)80439-8. [DOI] [PubMed] [Google Scholar]
  5. Goldman R. C., Sullivan P. A., Zakula D., Capobianco J. O. Kinetics of beta-1,3 glucan interaction at the donor and acceptor sites of the fungal glucosyltransferase encoded by the BGL2 gene. Eur J Biochem. 1995 Jan 15;227(1-2):372–378. doi: 10.1111/j.1432-1033.1995.tb20399.x. [DOI] [PubMed] [Google Scholar]
  6. Hartland R. P., Fontaine T., Debeaupuis J. P., Simenel C., Delepierre M., Latgé J. P. A novel beta-(1-3)-glucanosyltransferase from the cell wall of Aspergillus fumigatus. J Biol Chem. 1996 Oct 25;271(43):26843–26849. doi: 10.1074/jbc.271.43.26843. [DOI] [PubMed] [Google Scholar]
  7. Hearn V. M., Sietsma J. H. Chemical and immunological analysis of the Aspergillus fumigatus cell wall. Microbiology. 1994 Apr;140(Pt 4):789–795. doi: 10.1099/00221287-140-4-789. [DOI] [PubMed] [Google Scholar]
  8. Henrissat B., Bairoch A. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J. 1993 Aug 1;293(Pt 3):781–788. doi: 10.1042/bj2930781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henrissat B., Callebaut I., Fabrega S., Lehn P., Mornon J. P., Davies G. Conserved catalytic machinery and the prediction of a common fold for several families of glycosyl hydrolases. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7090–7094. doi: 10.1073/pnas.92.15.7090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Henrissat B., Davies G. Structural and sequence-based classification of glycoside hydrolases. Curr Opin Struct Biol. 1997 Oct;7(5):637–644. doi: 10.1016/s0959-440x(97)80072-3. [DOI] [PubMed] [Google Scholar]
  11. Jenkins J., Lo Leggio L., Harris G., Pickersgill R. Beta-glucosidase, beta-galactosidase, family A cellulases, family F xylanases and two barley glycanases form a superfamily of enzymes with 8-fold beta/alpha architecture and with two conserved glutamates near the carboxy-terminal ends of beta-strands four and seven. FEBS Lett. 1995 Apr 10;362(3):281–285. doi: 10.1016/0014-5793(95)00252-5. [DOI] [PubMed] [Google Scholar]
  12. Klis F. M. Review: cell wall assembly in yeast. Yeast. 1994 Jul;10(7):851–869. doi: 10.1002/yea.320100702. [DOI] [PubMed] [Google Scholar]
  13. Latgé J. P. Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev. 1999 Apr;12(2):310–350. doi: 10.1128/cmr.12.2.310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lemesle-Varloot L., Henrissat B., Gaboriaud C., Bissery V., Morgat A., Mornon J. P. Hydrophobic cluster analysis: procedures to derive structural and functional information from 2-D-representation of protein sequences. Biochimie. 1990 Aug;72(8):555–574. doi: 10.1016/0300-9084(90)90120-6. [DOI] [PubMed] [Google Scholar]
  15. MacLeod A. M., Tull D., Rupitz K., Warren R. A., Withers S. G. Mechanistic consequences of mutation of active site carboxylates in a retaining beta-1,4-glycanase from Cellulomonas fimi. Biochemistry. 1996 Oct 8;35(40):13165–13172. doi: 10.1021/bi9610616. [DOI] [PubMed] [Google Scholar]
  16. McCarter J. D., Withers S. G. Mechanisms of enzymatic glycoside hydrolysis. Curr Opin Struct Biol. 1994 Dec;4(6):885–892. doi: 10.1016/0959-440x(94)90271-2. [DOI] [PubMed] [Google Scholar]
  17. Mouyna I., Hartland R. P., Fontaine T., Diaquin M., Simenel C., Delepierre M., Henrissat B., Latgé J. P. A 1,3-beta-glucanosyltransferase isolated from the cell wall of Aspergillus fumigatus is a homologue of the yeast Bgl2p. Microbiology. 1998 Nov;144(Pt 11):3171–3180. doi: 10.1099/00221287-144-11-3171. [DOI] [PubMed] [Google Scholar]
  18. Mühlschlegel F. A., Fonzi W. A. PHR2 of Candida albicans encodes a functional homolog of the pH-regulated gene PHR1 with an inverted pattern of pH-dependent expression. Mol Cell Biol. 1997 Oct;17(10):5960–5967. doi: 10.1128/mcb.17.10.5960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakazawa T., Horiuchi H., Ohta A., Takagi M. Isolation and characterization of EPD1, an essential gene for pseudohyphal growth of a dimorphic yeast, Candida maltosa. J Bacteriol. 1998 Apr;180(8):2079–2086. doi: 10.1128/jb.180.8.2079-2086.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Popolo L., Vai M., Gatti E., Porello S., Bonfante P., Balestrini R., Alberghina L. Physiological analysis of mutants indicates involvement of the Saccharomyces cerevisiae GPI-anchored protein gp115 in morphogenesis and cell separation. J Bacteriol. 1993 Apr;175(7):1879–1885. doi: 10.1128/jb.175.7.1879-1885.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Popolo L., Vai M. The Gas1 glycoprotein, a putative wall polymer cross-linker. Biochim Biophys Acta. 1999 Jan 6;1426(2):385–400. doi: 10.1016/s0304-4165(98)00138-x. [DOI] [PubMed] [Google Scholar]
  22. Saporito-Irwin S. M., Birse C. E., Sypherd P. S., Fonzi W. A. PHR1, a pH-regulated gene of Candida albicans, is required for morphogenesis. Mol Cell Biol. 1995 Feb;15(2):601–613. doi: 10.1128/mcb.15.2.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vai M., Gatti E., Lacanà E., Popolo L., Alberghina L. Isolation and deduced amino acid sequence of the gene encoding gp115, a yeast glycophospholipid-anchored protein containing a serine-rich region. J Biol Chem. 1991 Jul 5;266(19):12242–12248. [PubMed] [Google Scholar]
  25. Woodcock S., Mornon J. P., Henrissat B. Detection of secondary structure elements in proteins by hydrophobic cluster analysis. Protein Eng. 1992 Oct;5(7):629–635. doi: 10.1093/protein/5.7.629. [DOI] [PubMed] [Google Scholar]

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