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

被引:98
作者
Briard, Jennie Grace [1 ]
Jiang, Hao [2 ,3 ,4 ]
Moremen, Kelley W. [5 ,6 ]
Macauley, Matthew Scott [1 ,7 ]
Wu, Peng [1 ]
机构
[1] Scripps Res Inst, Dept Mol Med, La Jolla, CA 92037 USA
[2] Ocean Univ China, Sch Med & Pharm, Minist Educ, Key Lab Marine Drugs, Qingdao 266003, Peoples R China
[3] Ocean Univ China, Sch Med & Pharm, Qingdao Natl Lab Marine Sci & Technol, Qingdao 266003, Peoples R China
[4] Ocean Univ China, Sch Med & Pharm, Shandong Prov Key Lab Glycosci & Glycoengn, Qingdao 266003, Peoples R China
[5] Univ Georgia, Complex Carbohydrate Res Ctr, 220 Riverbend Rd, Athens, GA 30602 USA
[6] Univ Georgia, Dept Biochem & Mol Biol, Athens, GA 30602 USA
[7] Univ Alberta, Dept Chem, Edmonton, AB T6G 2G2, Canada
关键词
LEWIS-X GLYCAN; SIALIC-ACID; OSTEOCLAST DIFFERENTIATION; CHEMOENZYMATIC SYNTHESIS; IMMUNE-SYSTEM; GLYCOSYLATION MUTANTS; SIGLEC LIGANDS; LIVING CELLS; IN-VIVO; LECTIN;
D O I
10.1038/s41467-018-03245-5
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
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页数:11
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