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Nat Commun. 2018 Feb 28;9(1):880. doi: 10.1038/s41467-018-03245-5.

Cell-based glycan arrays for probing glycan-glycan binding protein interactions.

Author information

1
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, 92037, USA.
2
Key Laboratory of Marine Drugs, Ministry of Education and Qingdao National Laboratory for Marine Science & Technology and Shandong Provincial Key Laboratory of Glycoscience & Glycoengineering, School of Medicine and Pharmacy, Ocean University of China, 266003, Qingdao, China.
3
Complex Carbohydrate Research Center and the Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
4
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, 92037, USA. macauley@ualberta.ca.
5
Department of Chemistry, University of Alberta, Edmonton, AB, T6G 2G2, Canada. macauley@ualberta.ca.
6
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, 92037, USA. pengwu@scripps.edu.

Abstract

Glycan microarrays provide a high-throughput means of profiling the interactions of glycan-binding proteins with their ligands. However, the construction of current glycan microarray platforms is time consuming and expensive. Here, we report a fast and cost-effective method for the assembly of cell-based glycan arrays to probe glycan-glycan-binding protein interactions directly on the cell surface. Chinese hamster ovary cell mutants with a narrow and relatively homogeneous repertoire of glycoforms serve as the foundation platforms to develop these arrays. Using recombinant glycosyltransferases, sialic acid, fucose, and analogs thereof are installed on cell-surface glycans to form cell-based arrays displaying diverse glycan epitopes that can be probed with glycan-binding proteins by flow cytometry. Using this platform, high-affinity glycan ligands are discovered for Siglec-15-a sialic acid-binding lectin involved in osteoclast differentiation. Incubating human osteoprogenitor cells with cells displaying a high-affinity Siglec-15 ligand impairs osteoclast differentiation, demonstrating the utility of this cell-based glycan array technology.

PMID:
29491407
PMCID:
PMC5830402
DOI:
10.1038/s41467-018-03245-5
[Indexed for MEDLINE]
Free PMC Article

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