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

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cell Differentiation
  • Cell Membrane / metabolism*
  • Cricetulus
  • Flow Cytometry
  • Fucose / analogs & derivatives
  • Fucose / metabolism
  • Glycosylation
  • Glycosyltransferases / metabolism
  • Humans
  • Immunoglobulins / metabolism*
  • Ligands
  • Membrane Proteins / metabolism*
  • Microscopy, Fluorescence
  • N-Acetylneuraminic Acid / analogs & derivatives
  • N-Acetylneuraminic Acid / metabolism
  • Osteoclasts / metabolism
  • Polysaccharides / metabolism*
  • Protein Array Analysis / methods*
  • Protein Binding
  • Recombinant Fusion Proteins / metabolism

Substances

  • Immunoglobulins
  • Ligands
  • Membrane Proteins
  • Polysaccharides
  • Recombinant Fusion Proteins
  • Siglec-15 protein, human
  • Fucose
  • Glycosyltransferases
  • N-Acetylneuraminic Acid