Silencing glycosaminoglycan functions in mouse embryonic stem cells with small molecule antagonists

Methods Enzymol. 2019:626:249-270. doi: 10.1016/bs.mie.2019.06.023. Epub 2019 Jul 24.

Abstract

Glycosylation is a ubiquitous post-translational modification that decorates proteins and lipids with glycans. These glycans can play critical roles in regulating biological events, and therefore, the discovery of strategies that target these molecules represent an important advancement toward understanding and controlling glycan-mediated cellular phenotypes. We describe the use of a small molecule, surfen, to temporarily silence the functions mediated by heparan sulfate glycosaminoglycans in mouse embryonic stem cells. Surfen binds heparan sulfate to antagonize growth factor interactions, thereby inhibiting signal transduction events that lead to differentiation. The strategies outlined in this chapter allow the characterization of resulting antagonistic effects caused by glycan-small molecule binding events toward maintaining embryonic stem cell pluripotency, curbing differentiation, and inhibiting signaling events.

Keywords: Glycans; Glycosaminoglycans; Glycosylation; Small molecule; Stem cells.

Publication types

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

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Electrophoresis, Polyacrylamide Gel / methods
  • Glycosaminoglycans / metabolism
  • Heparitin Sulfate / metabolism*
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Mice
  • Mouse Embryonic Stem Cells / cytology
  • Mouse Embryonic Stem Cells / drug effects*
  • Mouse Embryonic Stem Cells / metabolism
  • Polymerase Chain Reaction / methods
  • Signal Transduction / drug effects
  • Urea / analogs & derivatives*
  • Urea / pharmacology

Substances

  • Glycosaminoglycans
  • Intercellular Signaling Peptides and Proteins
  • aminoquinuride
  • Urea
  • Heparitin Sulfate