Engineered carbohydrate-recognition domains for glycoproteomic analysis of cell surface glycosylation and ligands for glycan-binding receptors

Methods Enzymol. 2010;480:165-79. doi: 10.1016/S0076-6879(10)80009-6.

Abstract

Modular calcium-dependent carbohydrate-recognition domains (CRDs) of mammalian glycan-binding receptors (C-type lectins), engineered to have novel glycan-binding selectivity, have been developed as tools for the study of glycans on cell surfaces. Structure-based specificity swapping between domains can be complemented by empirical characterization of ligand-binding specificity using glycan arrays. Both natural and modified CRDs can be used as probes for detecting and isolating glycoproteins that bear specific glycan epitopes and that act as target ligands for glycan-binding receptors. CRD-based affinity chromatography facilitates proteomic and glycomic analysis of such ligands.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Carbohydrate Metabolism* / physiology
  • Glycosylation
  • Humans
  • Lectins / chemistry
  • Lectins / metabolism*
  • Ligands
  • Membrane Glycoproteins / analysis
  • Membrane Glycoproteins / chemical synthesis*
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Binding
  • Protein Engineering / methods*
  • Protein Interaction Domains and Motifs*
  • Proteomics / methods*

Substances

  • Lectins
  • Ligands
  • Membrane Glycoproteins