Using a Modified Proximity Ligation Protocol to Study the Interaction Between Chaperones and Associated Proteins

Simone Baldan, Anatoli B. Meriin, Michael Y. Sherman

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

Abstract

Molecular chaperones can interact with multiple proteins to form large networks. Understanding these interactions may shed light on the complexity of the chaperone functions. Here we developed a protocol for a modified proximity ligation-based methodology (PLA) for the detection of protein–protein interactions in order to understand how the Hsp70-Bag3 complex interacts with components of the Hippo signaling pathway. These experiments helped to elucidate the mechanisms of transmission of the proteotoxic stress signal to the Hippo pathway. The modified PLA technology has many advantages compared to co-immunoprecipitation protocols. It has higher sensitivity, is quantitative, and can be done in a 96-well format.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages163-174
Number of pages12
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2693
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Bag3
  • Hippo pathway
  • Hsp70
  • LATS1/2
  • Protein complex
  • Proximity ligation assay
  • Yap

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