Unravelling glucan recognition systems by glycome microarrays using the designer approach and mass spectrometry.

Autor:	Palma AS; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom; §UCIBIO-REQUIMTE, Department of Chemistry, Faculty of Science and Technology, NOVA University of Lisbon; angelina.palma@fct.unl.pt w.chai@imperial.ac.uk., Liu Y; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom;, Zhang H; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom;, Zhang Y; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom;, McCleary BV; ¶Megazyme International, County Wicklow, Ireland;, Yu G; ‖Key Laboratory of Marine Drugs, Shandong Provincial Key Laboratory of Glycoscience and Glycotechnology, School of Medicine and Pharmacy, Ocean University of China;, Huang Q; *Department of Chemistry, Wuhan University, Wuhan, China;, Guidolin LS; ‡‡Instituto de Investigaciones Biotecnológicas 'Dr. Rodolfo A. Ugalde', Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín, Buenos Aires, Argentina;, Ciocchini AE; ‡‡Instituto de Investigaciones Biotecnológicas 'Dr. Rodolfo A. Ugalde', Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín, Buenos Aires, Argentina;, Torosantucci A; §§Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanitá, Rome, Italy;, Wang D; ¶¶Tumor Glycomics Laboratory, SRI International Biosciences Division, Menlo Park, CA;, Carvalho AL; §UCIBIO-REQUIMTE, Department of Chemistry, Faculty of Science and Technology, NOVA University of Lisbon;, Fontes CM; ‖‖CIISA, Faculdade de Medicina Veterinária, Universidade de Lisboa, Portugal., Mulloy B; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom;, Childs RA; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom;, Feizi T; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom;, Chai W; From the ‡Glycosciences Laboratory, Department of Medicine, Imperial College London, United Kingdom; angelina.palma@fct.unl.pt w.chai@imperial.ac.uk.
Jazyk:	angličtina
Zdroj:	Molecular & cellular proteomics : MCP [Mol Cell Proteomics] 2015 Apr; Vol. 14 (4), pp. 974-88. Date of Electronic Publication: 2015 Feb 10.
DOI:	10.1074/mcp.M115.048272
Abstrakt:	Glucans are polymers of d-glucose with differing linkages in linear or branched sequences. They are constituents of microbial and plant cell-walls and involved in important bio-recognition processes, including immunomodulation, anticancer activities, pathogen virulence, and plant cell-wall biodegradation. Translational possibilities for these activities in medicine and biotechnology are considerable. High-throughput micro-methods are needed to screen proteins for recognition of specific glucan sequences as a lead to structure-function studies and their exploitation. We describe construction of a "glucome" microarray, the first sequence-defined glycome-scale microarray, using a "designer" approach from targeted ligand-bearing glucans in conjunction with a novel high-sensitivity mass spectrometric sequencing method, as a screening tool to assign glucan recognition motifs. The glucome microarray comprises 153 oligosaccharide probes with high purity, representing major sequences in glucans. Negative-ion electrospray tandem mass spectrometry with collision-induced dissociation was used for complete linkage analysis of gluco-oligosaccharides in linear "homo" and "hetero" and branched sequences. The system is validated using antibodies and carbohydrate-binding modules known to target α- or β-glucans in different biological contexts, extending knowledge on their specificities, and applied to reveal new information on glucan recognition by two signaling molecules of the immune system against pathogens: Dectin-1 and DC-SIGN. The sequencing of the glucan oligosaccharides by the MS method and their interrogation on the microarrays provides detailed information on linkage, sequence and chain length requirements of glucan-recognizing proteins, and are a sensitive means of revealing unsuspected sequences in the polysaccharides. (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
Databáze:	MEDLINE
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