Metabolic labeling of glycoconjugates with photocrosslinking sugars

Methods Enzymol. 2010:478:541-62. doi: 10.1016/S0076-6879(10)78026-5.

Abstract

Protein-carbohydrate interactions play essential roles in a variety of biological processes. This class of interactions is particularly important in development, immunology, infection, and carcinogenesis. However, the transient nature of glycan-dependent interactions hampers efforts to detect and characterize these complexes. Photocrosslinking is emerging as a powerful tool to discover and study glycan-dependent complexes. Through the use of photocrosslinking groups, UV irradiation can be employed to introduce a covalent bond between two transiently interacting molecules. Here we describe the use of metabolic oligosaccharide engineering to incorporate a photocrosslinkable sugar into cellular glycoconjugates and the use of this photocrosslinker to covalently capture glycan-mediated interactions.

Publication types

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

MeSH terms

  • Carbohydrate Sequence
  • Cells, Cultured
  • Cross-Linking Reagents / chemistry*
  • Glycoconjugates / chemistry*
  • Humans
  • Jurkat Cells
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / metabolism
  • Molecular Sequence Data
  • N-Acetylneuraminic Acid / chemistry
  • Photoaffinity Labels* / chemistry
  • Photoaffinity Labels* / metabolism
  • Polysaccharides / chemistry*
  • Protein Engineering / methods

Substances

  • Cross-Linking Reagents
  • Glycoconjugates
  • Membrane Glycoproteins
  • Photoaffinity Labels
  • Polysaccharides
  • N-Acetylneuraminic Acid