Neuronal activation by GPI-linked neuroligin-1 displayed in synthetic lipid bilayer membranes

Langmuir. 2005 Nov 8;21(23):10693-8. doi: 10.1021/la051243d.

Abstract

We have characterized, in vitro, interactions between hippocampal neuronal cells and silica microbeads coated with synthetic, fluid, lipid bilayer membranes containing the glycosylphosphatidyl inositol (GPI)-linked extracellular domain of the postsynaptic membrane protein neuroligin-1. These bilayer-neuroligin-1 beads activated neuronal cells to form presynaptic nerve terminals at the point of contact in a manner similar to that observed for live PC12 cells, ectopically expressing the full length neuroligin-1. The synthetic membranes exhibited biological activity at neuroligin-1 densities of approximately 1 to 6 proteins/microm(2). Polyolycarbonate beads with neuroligin-1 covalently attached to the surface failed to activate neurons despite the fact that neuroligin-1 binding activity is preserved. This implies that a lipid membrane environment is likely to be essential for neuroligin-1 activity. This technique allows the study of isolated proteins in an environment that has physical properties resembling those of a cell surface; proteins can diffuse freely within the membrane, retain their in vivo orientations, and are in a nondenatured state. In addition, the synthetic membrane environment affords control over both lipid and protein composition. This technology is easily implemented and can be applied to a wide variety of cellular studies.

Publication types

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

MeSH terms

  • Animals
  • Cell Adhesion Molecules, Neuronal
  • Cell Line
  • Glycosylphosphatidylinositols / metabolism*
  • Humans
  • Lipid Bilayers*
  • Membrane Proteins / metabolism*
  • Nerve Tissue Proteins / metabolism*
  • Neurons / metabolism*
  • Rats

Substances

  • Cell Adhesion Molecules, Neuronal
  • Glycosylphosphatidylinositols
  • Lipid Bilayers
  • Membrane Proteins
  • Nerve Tissue Proteins
  • neuroligin 1