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

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

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 id...

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Bibliographic Details
Published in:Biochemical journal Vol. 457; no. 2; p. 347
Main Authors: Fabre, Emeline, Sfihi-Loualia, Ghenima, Pourcelot, Marilyne, Coddeville, Bernadette, Krzewinski, Frédéric, Bouckaert, Julie, Maes, Emmanuel, Hurtaux, Thomas, Dubois, Romaric, Fradin, Chantal, Mallet, Jean-Maurice, Poulain, Daniel, Delplace, Florence, Guerardel, Yann
Format: Journal Article
Language:English
Published: England 15-01-2014
Subjects:
Candida albicans - enzymology
Candida albicans - genetics
Cell Wall - enzymology
Mannans - chemistry
Mannans - genetics
Mannans - metabolism
Mannose - chemistry
Mannose - genetics
Mannose - metabolism
Mannosyltransferases - chemistry
Mannosyltransferases - genetics
Mannosyltransferases - metabolism
Phosphopeptides - chemistry
Phosphopeptides - genetics
Phosphopeptides - metabolism
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
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Summary: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.
ISSN:1470-8728
DOI:10.1042/BJ20131012