Authors: Zhang, Hongtao; Palma, Angelina S.; Zhang, Yibing; Childs, Robert A.; Liu, Yan; Mitchell, Daniel A.; Guidolin, Leticia Soledad; Weigel, Wilfried; Mulloy, Barbara; Ciocchini, Andres Eduardo; Feizi, Ten; Chai, Wengang
Publication Date: 2016.
Language: English.
Abstract:
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 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 β1,2-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 β1,2-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 CβG by this receptor. These findings open the way to unravelling mechanisms of immunomodulation mediated by β1,2-glucans in mammalian systems.
Author affiliation: Zhang, Hongtao. Imperial College London; Reino Unido. Jiangnan University; China
Author affiliation: Palma, Angelina S.. Imperial College London; Reino Unido. Universidade de Lisboa; Portugal
Author affiliation: Zhang, Yibing. Imperial College London; Reino Unido
Author affiliation: Childs, Robert A.. Imperial College London; Reino Unido
Author affiliation: Liu, Yan. Imperial College London; Reino Unido
Author affiliation: Mitchell, Daniel A.. University of Warwick; Reino Unido
Author affiliation: Guidolin, Leticia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; Argentina
Author affiliation: Weigel, Wilfried. SCIENION AG; Alemania
Author affiliation: Mulloy, Barbara. Imperial College London; Reino Unido
Author affiliation: Ciocchini, Andres Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; Argentina
Author affiliation: Feizi, Ten. Imperial College London; Reino Unido
Author affiliation: Chai, Wengang. Imperial College London; Reino Unido
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Palma, Angelina S.; Liu, Yan; Zhang, Hongtao; Zhang, Yibing; McCleary, Barry V.; Yu, Guangli; Huang, Qilin; Guidolin, Leticia Soledad; Ciocchini, Andres Eduardo; Torosantucci, Antonella; Wang, Denong; Carvalho, Ana Luísa; Fontes, Carlos M. G. A.; Mulloy, Barbara; Childs, Robert A.; Feizi, Ten; Chai, Wengang
Publication Date: 2015.
Language: English.
Abstract:
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 cellwall 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.
Author affiliation: Palma, Angelina S.. Imperial College London; Reino Unido. Universidade de Lisboa; Portugal
Author affiliation: Liu, Yan. Imperial College London; Reino Unido
Author affiliation: Zhang, Hongtao. Imperial College London; Reino Unido. Jiangnan University; China
Author affiliation: Zhang, Yibing. Imperial College London; Reino Unido
Author affiliation: McCleary, Barry V.. Megazyme International Ireland; Irlanda
Author affiliation: Yu, Guangli. Ocean University Of China; China
Author affiliation: Huang, Qilin. Huazhong Agricultural University; China. Wuhan University; China
Author affiliation: Guidolin, Leticia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Author affiliation: Ciocchini, Andres Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Author affiliation: Torosantucci, Antonella. Istituto Superiore Di Sanita; Italia
Author affiliation: Wang, Denong. Sri International; Estados Unidos
Author affiliation: Carvalho, Ana Luísa. Universidade de Lisboa; Portugal
Author affiliation: Fontes, Carlos M. G. A.. Universidade de Lisboa; Portugal
Author affiliation: Mulloy, Barbara. Imperial College London; Reino Unido
Author affiliation: Childs, Robert A.. Imperial College London; Reino Unido
Author affiliation: Feizi, Ten. Imperial College London; Reino Unido
Author affiliation: Chai, Wengang. Imperial College London; Reino Unido
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas