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Journal of Biochemistry Advance Access published online on January 17, 2009
Journal of Biochemistry, 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, France.
2Unité de Physiopathologie des Candidoses, Institut National de la Santé et de la Recherche Médicale (Inserm) U799, Université Lille 2, 59045, Lille, France.
3Institut Pasteur, Unité de Mycologie Moléculaire ; CNRS, URA3012, F-75015, France

*To whom correspondence should be addressed: Yann Guérardel: tel : +33(0)320336347 Fax : +33(0)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


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