Membrane display and functional analysis of juxtacrine ligand-receptor signaling

Biotechniques. 2015 Oct 1;59(4):231-8, 240. doi: 10.2144/000114342. eCollection 2015 Oct.

Abstract

We developed a strategy for identifying modulators of juxtacrine signaling, triggered by a cell-surface ligand displayed on synthetic lipid bilayers, via cognate receptors on apposed cells. Using readouts for receptor lateral transport and intracellular signaling, we screened a small interfering RNA (siRNA) library and identified specific receptor tyrosine kinases (RTKs) that directly or indirectly modulate apoptosis signaling by a model death ligand through its cognate death receptors. This approach may be broadly useful for studying juxtacrine cell-cell signaling systems.

Keywords: apoptosis; caspase; death receptor; juxtacrine; screen; supported membrane display; transport.

MeSH terms

  • Apoptosis / genetics*
  • Cell Communication / genetics*
  • Cell Line, Tumor
  • Gene Expression Regulation
  • Humans
  • Ligands
  • Lipid Bilayers / chemical synthesis
  • Lipid Bilayers / chemistry
  • Lipid Bilayers / metabolism
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / genetics
  • Receptors, Death Domain / biosynthesis
  • Receptors, Death Domain / genetics
  • Signal Transduction / genetics
  • TNF-Related Apoptosis-Inducing Ligand / chemistry
  • TNF-Related Apoptosis-Inducing Ligand / genetics
  • TNF-Related Apoptosis-Inducing Ligand / metabolism*

Substances

  • Ligands
  • Lipid Bilayers
  • RNA, Small Interfering
  • Receptors, Death Domain
  • TNF-Related Apoptosis-Inducing Ligand