Dataset Information

Generation and characterization of ?1,2-gluco-oligosaccharide probes from Brucella abortus cyclic ?-glucan and their recognition by C-type lectins of the immune system.

ABSTRACT: The ?1,2-glucans produced by bacteria are important in invasion, survival and immunomodulation in infected hosts be they mammals or plants. However, there has been a lack of information on proteins which recognize these molecules. This is partly due to the extremely limited availability of the sequence-defined oligosaccharides and derived probes for use in the study of their interactions. Here we have used the cyclic ?1,2-glucan (C?G) of the bacterial pathogen Brucella abortus, after removal of succinyl side chains, to prepare linearized oligosaccharides which were used to generate microarrays. We describe optimized conditions for partial depolymerization of the cyclic glucan by acid hydrolysis and conversion of the ?1,2-gluco-oligosaccharides, with degrees of polymerization 2-13, to neoglycolipids for the purpose of generating microarrays. By microarray analyses, we show that the C-type lectin receptor DC-SIGNR, like the closely related DC-SIGN we investigated earlier, binds to the ?1,2-gluco-oligosaccharides, as does the soluble immune effector serum mannose-binding protein. Exploratory studies with DC-SIGN are suggestive of the recognition also of the intact C?G by this receptor. These findings open the way to unravelling mechanisms of immunomodulation mediated by ?1,2-glucans in mammalian systems.

SUBMITTER: Zhang H

PROVIDER: S-EPMC5072146 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature

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Publications

Generation and characterization of β1,2-gluco-oligosaccharide probes from Brucella abortus cyclic β-glucan and their recognition by C-type lectins of the immune system.

Zhang Hongtao H Palma Angelina S AS Zhang Yibing Y Childs Robert A RA Liu Yan Y Mitchell Daniel A DA Guidolin Leticia S LS Weigel Wilfried W Mulloy Barbara B Ciocchini Andrés E AE Feizi Ten T Chai Wengang W

Glycobiology 20160406 10

The β1,2-glucans produced by bacteria are important in invasion, survival and immunomodulation in infected hosts be they mammals or plants. However, there has been a lack of information on proteins which recognize these molecules. This is partly due to the extremely limited availability of the sequence-defined oligosaccharides and derived probes for use in the study of their interactions. Here we have used the cyclic β1,2-glucan (CβG) of the bacterial pathogen Brucella abortus, after removal of ...[more]

PMID: 27053576

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Project description:UnlabelledCyclic ?-1,2-glucans (C?G) are periplasmic homopolysaccharides that play an important role in the virulence and interaction of Brucella with the host. Once synthesized in the cytoplasm by the C?G synthase (Cgs), C?G are transported to the periplasm by the C?G transporter (Cgt) and succinylated by the C?G modifier enzyme (Cgm). Here, we used a bacterial two-hybrid system and coimmunoprecipitation techniques to study the interaction network between these three integral inner membrane proteins. Our results indicate that Cgs, Cgt, and Cgm can form both homotypic and heterotypic interactions. Analyses carried out with Cgs mutants revealed that the N-terminal region of the protein (Cgs region 1 to 418) is required to sustain the interactions with Cgt and Cgm as well as with itself. We demonstrated by single-cell fluorescence analysis that in Brucella, Cgs and Cgt are focally distributed in the membrane, particularly at the cell poles, whereas Cgm is mostly distributed throughout the membrane with a slight accumulation at the poles colocalizing with the other partners. In summary, our results demonstrate that Cgs, Cgt, and Cgm form a membrane-associated biosynthetic complex. We propose that the formation of a membrane complex could serve as a mechanism to ensure the fidelity of C?G biosynthesis by coordinating their synthesis with the transport and modification.ImportanceIn this study, we analyzed the interaction and localization of the proteins involved in the synthesis, transport, and modification of Brucella abortus cyclic ?-1,2-glucans (C?G), which play an important role in the virulence and interaction of Brucella with the host. We demonstrate that these proteins interact, forming a complex located mainly at the cell poles; this is the first experimental evidence of the existence of a multienzymatic complex involved in the metabolism of osmoregulated periplasmic glucans in bacteria and argues for another example of pole differentiation in Brucella. We propose that the formation of this membrane complex could serve as a mechanism to ensure the fidelity of C?G biosynthesis by coordinating synthesis with the transport and modification.

Dataset Information

Generation and characterization of ?1,2-gluco-oligosaccharide probes from Brucella abortus cyclic ?-glucan and their recognition by C-type lectins of the immune system.

Publications

Generation and characterization of β1,2-gluco-oligosaccharide probes from Brucella abortus cyclic β-glucan and their recognition by C-type lectins of the immune system.

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