Protocol for visualizing conditional interaction between transmembrane and cytoplasmic proteins

STAR Protoc. 2021 Mar 31;2(2):100430. doi: 10.1016/j.xpro.2021.100430. eCollection 2021 Jun 18.

Abstract

This protocol visualizes dynamic interaction between a transmembrane protein and an intracellular protein induced by clusterization/oligomerization of the transmembrane protein. Association-dissociation of the intracellular region of the transmembrane protein with cytoplasmic protein(s) is detected by proximity ligation assay. Since a transmembrane protein often resists extraction, biochemical analysis of its dynamic interaction with cytoplasmic effectors is cumbersome. This protocol quantitatively visualizes protein-protein interaction occurring in the membrane periphery, providing a powerful tool to elucidate signal transduction across the membrane. For complete details on the use and execution of this protocol, please refer to Ooki et al. (2019).

Keywords: Cell Biology; Cell Membrane; Cell culture; Microscopy; Protein Biochemistry; Signal Transduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cytoplasm* / chemistry
  • Cytoplasm* / metabolism
  • Humans
  • Membrane Proteins* / analysis
  • Membrane Proteins* / chemistry
  • Membrane Proteins* / metabolism
  • Molecular Imaging / methods*
  • Molecular Probes
  • Protein Binding / physiology
  • Signal Transduction / physiology

Substances

  • Membrane Proteins
  • Molecular Probes