Protein micropatterning assay: Quantitative analysis of protein–protein interactions

Gerhard Schütz, Julian Weghuber, Peter Lanzerstorfer, Eva Sevcsik

Research output: Chapter in Book/Report/Conference proceedingsChapterpeer-review

5 Citations (Scopus)

Abstract

Characterization, especially quantification, of protein interactions in live cells is usually not an easy endeavor. Here, we describe a straightforward method to identify and quantify the interaction of a membrane protein (“bait”) and a fluorescently labeled interaction partner (“prey”) (membrane-bound or cyto-solic) in live cells using Total Internal Reflection Fluorescence microscopy. The bait protein is immobilized within patterns in the plasma membrane (e.g., via an antibody); the bait–prey interaction strength can be quantified by determining the prey bulk fluorescence intensity with respect to the bait patterns. This method is particularly suitable also for the analysis of weak, transient interactions that are not easily accessible with other methods.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages261-270
Number of pages10
Volume1550
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1550
ISSN (Print)1064-3745

Keywords

  • Membrane proteins
  • Micropatterning
  • Protein–protein interactions
  • Quantitive analysis
  • Soft lithography
  • TIRF microscopy
  • Proteins/metabolism
  • Protein Interaction Mapping/methods
  • Workflow
  • Membrane Proteins/metabolism
  • Fluorescent Antibody Technique
  • Protein Binding
  • Microscopy, Fluorescence

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