Analysis of CYLD proteolysis by CASPASE 8 in bone marrow-derived macrophages

Diana Legarda, Adrian T. Ting

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Previous studies have demonstrated that CASPASE 8 can generate a prosurvival signal by inhibiting necroptosis via the cleavage of the deubiquitinating enzyme CYLD. Cleavage of CYLD at D215 results in the generation of a 25 kD N-terminal fragment and degradation of the C-terminal fragment containing the catalytic domain. Since CYLD is required for TNF-induced necroptosis, its proteolysis is necessary and sufficient to suppress necroptosis and generate a survival signal. Here we describe how to visualize CYLD proteolysis by western blot analysis, as a measure of CASPASE 8 activity and inhibition of necroptosis.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages181-188
Number of pages8
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1857
ISSN (Print)1064-3745

Keywords

  • Bone marrow-derived macrophages
  • Cleavage
  • CYLD
  • Necroptosis
  • Proteolysis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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