Liposomal Permeabilization Assay to Study the Functional Interactions of the BCL-2 Family

Denis E. Reyna, Evripidis Gavathiotis

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Apoptosis, a form of programmed cell death that is important for development and homeostasis, is regulated by the BCL-2 family of proteins. Over twenty BCL-2 family members have been classified in three groups based on structural homology and function. The multidomain antiapoptotic proteins promote survival, whereas the multidomain and the BH3-only proapoptotic members induce cell death. Because the interaction among the BCL-2 family members occurs primarily at the mitochondrial outer membrane, biochemical assays using artificial liposomes have been developed to study the functional relationship between these proteins. The liposomal permeabilization assay is a cell-free system that relies on the ability of multidomain pro-apoptotic members to promote membrane permeabilization upon activation. By encapsulating a fluorophore and a quencher into liposomes, the degree of permeabilization can be quantified by the increase in fluorescence intensity as the fluorophore and quencher dissociate. The liposomal permeabilization assay has been used to delineate interactions among BCL-2 family members as well as to characterize peptides, small molecules, and lipids that modulate the function of BCL-2 family of proteins. Here, we describe in detail the permeabilization of liposomes induced by the interaction between BAX and BH3-only activator tBID.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages111-119
Number of pages9
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1877
ISSN (Print)1064-3745

Keywords

  • Apoptosis
  • BAK
  • BAX
  • BCL-2 family
  • BH3-domain
  • Liposomal membrane
  • Mitochondria
  • MOMP
  • tBID

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Reyna, D. E., & Gavathiotis, E. (2019). Liposomal Permeabilization Assay to Study the Functional Interactions of the BCL-2 Family. In Methods in Molecular Biology (pp. 111-119). (Methods in Molecular Biology; Vol. 1877). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8861-7_7