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Study of GPCR–Protein Interactions Using Gel Overlay Assays and Glutathione-S-Transferase-Fusion Protein Pull-Downs

  • Ashley E. Brady
  • Yunjia Chen
  • Lee E. Limbird
  • Qin WangEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 746)

Abstract

Numerous recent studies have suggested that the predicted cytosolic domains of G protein-coupled receptors represent a surface for association with proteins that may serve multiple roles in receptor localization, turnover, and signaling beyond the well-characterized interactions of these receptors with heterotrimeric G proteins. This Chapter describes two in vitro methods for ascertaining interactions between G protein-coupled receptors and various binding partners: gel overlay strategies and GST-fusion protein pull-downs.

Key words

Gel overlay G protein-coupled receptor Glutathione-S-transferase pull-down Protein–protein interaction 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ashley E. Brady
  • Yunjia Chen
  • Lee E. Limbird
  • Qin Wang
    • 1
    Email author
  1. 1.Department of Physiology and BiophysicsUniversity of Alabama at BirminghamBirminghamUSA

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