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Characterization of the recombinant Candida albicans β-1,2-mannosyltransferase that initiates the β-mannosylation of cell wall phosphopeptidomannan

Authors :
Ghenima Sfihi-Loualia
Bernadette Coddeville
Julie Bouckaert
Thomas Hurtaux
Jean-Maurice Mallet
Frédéric Krzewinski
Yann Guérardel
Emeline Fabre
Florence Delplace
Daniel Poulain
Chantal Fradin
Romaric Dubois
Emmanuel Maes
Marilyne Pourcelot
Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF)
Université de Lille-Centre National de la Recherche Scientifique (CNRS)
Université Pierre et Marie Curie - Paris 6 (UPMC)
Physiopathologie des Candidoses
Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Droit et Santé
Inflammation: mécanismes et régulation et interactions avec la nutrition et les candidoses
Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)
Université de Lille, Droit et Santé-Institut National de la Santé et de la Recherche Médicale (INSERM)
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF)
Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)
Source :
Biochemical Journal, Biochemical Journal, 2013, 457 (2), pp.347-360. ⟨10.1042/BJ20131012⟩, Biochemical Journal, Portland Press, 2013, 457 (2), pp.347-360. ⟨10.1042/BJ20131012⟩
Publication Year :
2013
Publisher :
Portland Press Ltd., 2013.

Abstract

The presence of β-mannosides in their cell walls confers specific features on the pathogenic yeasts Candida albicans and Candida glabrata compared with non-pathogenic yeasts. In the present study, we investigated the enzymatic properties of Bmt1 (β-mannosyltransferase 1), a member of the recently identified β-mannosyltransferase family, from C. albicans. A recombinant soluble enzyme lacking the N-terminal region was expressed as a secreted protein from the methylotrophic yeast Pichia pastoris. In parallel, functionalized natural oligosaccharides isolated from Saccharomyces cerevisiae and a C. albicans mutant strain, as well as synthetic α-oligomannosides, were prepared and used as potential acceptor substrates. Bmt1p preferentially utilizes substrates containing linear chains of α-1,2-linked mannotriose or mannotetraose. The recombinant enzyme consecuti-vely transfers two mannosyl units on to these acceptors, leading to the production of α-mannosidase-resistant oligomannosides. NMR experiments further confirmed the presence of a terminal βMan (β-1,2-linked mannose) unit in the first enzyme product. In the future, a better understanding of specific β-1,2-mannosyltransferase molecular requirements will help the design of new potential antifungal drugs.

Details

ISSN :
14708728 and 02646021
Volume :
457
Database :
OpenAIRE
Journal :
Biochemical Journal
Accession number :
edsair.doi.dedup.....a6d6d9b7fa03ecf469cb9c8bb125d32d
Full Text :
https://doi.org/10.1042/bj20131012