Measuring ER stress and the unfolded protein response using mammalian tissue culture system

Methods Enzymol. 2011:490:71-92. doi: 10.1016/B978-0-12-385114-7.00004-0.

Abstract

The endoplasmic reticulum (ER) functions to properly fold and process secreted and transmembrane proteins. Environmental and genetic factors that disrupt ER function cause an accumulation of misfolded and unfolded proteins in the ER lumen, a condition termed ER stress. ER stress activates a signaling network called the Unfolded Protein Response (UPR) to alleviate this stress and restore ER homeostasis, promoting cell survival and adaptation. However, under unresolvable ER stress conditions, the UPR promotes apoptosis. Here, we discuss the current methods to measure ER stress levels, UPR activation, and subsequent pathways in mammalian cells. These methods will assist us in understanding the UPR and its contribution to ER stress-related disorders such as diabetes and neurodegeneration.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activating Transcription Factor 6 / genetics
  • Activating Transcription Factor 6 / metabolism
  • Animals
  • Apoptosis
  • Biomarkers / metabolism
  • Cell Survival
  • Cells, Cultured
  • Endoplasmic Reticulum / pathology
  • Endoplasmic Reticulum / physiology*
  • Endoplasmic Reticulum / ultrastructure
  • Enzyme Activation
  • Homeostasis
  • Humans
  • Protein Folding
  • RNA, Messenger / metabolism
  • Signal Transduction / physiology
  • Stress, Physiological*
  • Tissue Culture Techniques / instrumentation
  • Tissue Culture Techniques / methods*
  • Transcriptional Activation
  • Unfolded Protein Response / physiology*
  • eIF-2 Kinase / genetics
  • eIF-2 Kinase / metabolism

Substances

  • Activating Transcription Factor 6
  • Biomarkers
  • RNA, Messenger
  • PERK kinase
  • eIF-2 Kinase