Methods to Study the Roles of β-Arrestins in Meningococcal Signaling

Methods Mol Biol. 2019:1957:325-334. doi: 10.1007/978-1-4939-9158-7_20.

Abstract

Neisseria meningitidis is a Gram-negative diplococcus restricted to humans that causes severe septicemia and/or meningitidis. Initial adhesion to human endothelial cells is mediated through the interaction of type IV pili with the hetero-oligomeric complexes formed by the human receptors CD147 and the β2-adrenergic receptor. Interaction with this complex heterodimer activates a β-arrestin-biased signaling pathway leading to actin polymerization and accumulation of ezrin and ezrin-binding partners. These signaling events promote the formation of cell plasma membrane protrusions in endothelial cells, which are crucial for N. meningitidis colonies to resist shear stress and colonize blood vessels. Here we provide detailed protocols to evaluate the role of β-arrestins in actin and ezrin signaling downstream of G protein-coupled receptor activation.

Keywords: Membrane protrusion; Neisseria meningitidis; β-Arrestins; β2-Adrenergic receptor.

MeSH terms

  • Cells, Cultured
  • Endothelial Cells / metabolism
  • Endothelial Cells / microbiology
  • HEK293 Cells
  • Humans
  • Molecular Biology / methods*
  • Neisseria meningitidis / metabolism*
  • RNA, Small Interfering / metabolism
  • Receptors, G-Protein-Coupled / metabolism
  • Signal Transduction*
  • beta-Arrestins / metabolism*

Substances

  • RNA, Small Interfering
  • Receptors, G-Protein-Coupled
  • beta-Arrestins