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

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 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Biochemistry Oxford University Press

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

Abstract

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
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