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Journal of Biochemistry Advance Access originally published online on January 17, 2009
Journal of Biochemistry 2009 145(4):413-419; doi:10.1093/jb/mvp008
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© The Authors 2009. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved

Rapid Communication

Molecular Phenotyping of Mannosyltransferases-Deficient Candida albicans Cells by High-Resolution Magic Angle Spinning NMR

Emmanuel Maes1, Céline Mille2, Xavier Trivelli1, Guilhem Janbon3, Daniel Poulain2 and Yann Guérardel1,*

1Unité de Glycobiologie Structurale et Fonctionnelle, UMR CNRS 8576, IFR 147, Université des Sciences et Technologies de Lille 1, 59655, Villeneuve d’Ascq; 2Unité de Physiopathologie des Candidoses, Institut National de la Santé et de la Recherche Médicale (Inserm) U799, Université Lille 2, 59045, Lille; and 3Institut Pasteur, Unité de Mycologie Moléculaire; CNRS, URA3012, F-75015, France

*To whom correspondence should be addressed. Tel: +33-320336347, Fax: +33-320436555, E-mail: yann.guerardel{at}univ-lille1.fr

Received December 11, 2008; Accepted December 26, 2008


  Abstract

The yeast Candida albicans is an opportunistic pathogen that causes infections in immunocompromised individuals with a high morbidity and mortality levels. Recognition of yeasts by host cells is directly mediated by cell wall components of the yeast, including a wide range of abundantly expressed glycoconjugates. Of particular interest in C. albicans are the β-mannosylated epitopes that show a complex expression pattern on N-glycan moiety of phosphopeptidomannans and are absent in the non-pathogenic species Saccharomyces cerevisiae. Being known as potent antigens for the adaptive immune response and elicitors of specific infection-protective antibodies, the exact delineation of β-mannosides regulation and expression pathways has lately become a major milestone toward the comprehension of host-pathogen interplay. Using the newly developed HR-MAS NMR methodology, we demonstrate the possibility of assessing the general profiles of cell-surface-exposed glycoconjugates from intact living yeast cells without any prior purification step. This technique permitted to directly observe structural modifications of surface expressed phosphodiester-linked β-mannosides on a series of deletion strains in β-mannosyltransferases and phospho-mannosyltransferases compared with their parental strains

Key Words: Candida albicans, high-resolution magic angle NMR, β-mannosylation, mannosyltransferase

Abbreviations: HR-MAS NMR, high-resolution magic angle spinning nuclear magnetic resonance; Man, mannose; Manp, mannopyranoside, GlcpNAc, N-acetylglucopyranoside, PPM, phosphopeptidomannan; HSQC, heteronuclear single quantum coherence; BMT, β-mannosyltransferase


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