Methods to Study the BECN1 Interactome in the Course of Autophagic Responses

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Autophagy is an extremely dynamic process that mediates the rapid degradation of intracellular components in response to different stress conditions. The autophagic response is executed by specific protein complexes, whose function is regulated by posttranslational modifications and interactions with positive and negative regulators. A comprehensive analysis of how autophagy complexes are temporally modified upon stress stimuli is therefore particularly relevant to understand how this pathway is regulated. Here, we describe a method to define the protein–protein interaction network of a central complex involved in autophagy induction, the Beclin 1 complex. This method is based on the quantitative comparison of protein complexes immunopurified at different time points using a stable isotope labeling by amino acids in cell culture approach. Understanding how the Beclin 1 complex dynamically changes in response to different stress stimuli may provide useful insights to disclose novel molecular mechanisms responsible for the dysregulation of autophagy in pathological conditions, such as cancer, neurodegeneration, and infections.

Original languageEnglish
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages429-445
Number of pages17
Volume587
DOIs
Publication statusPublished - Jan 1 2017

Publication series

NameMethods in Enzymology
Volume587
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Autophagic response
  • Beclin 1 complex
  • Immunoprecipitation
  • Interactome
  • Mass spectrometry
  • Protein complexes
  • Proteomics
  • SILAC

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'Methods to Study the BECN1 Interactome in the Course of Autophagic Responses'. Together they form a unique fingerprint.

  • Cite this

    Antonioli, M., Ciccosanti, F., Dengjel, J., & Fimia, G. M. (2017). Methods to Study the BECN1 Interactome in the Course of Autophagic Responses. In Methods in Enzymology (Vol. 587, pp. 429-445). (Methods in Enzymology; Vol. 587). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2016.09.069