• Assay Selection Tool

BellBrook Labs

  • Products
    • Transcreener® HTS Assay Kits
      • Transcreener® ADP² Kinase Assay Kits
        • Transcreener® ADP2 Assay Kit – FP Readout
        • Transcreener® ADP2 Assay Kit – FI Readout
        • Transcreener® ADP2 Assay Kit – TR-FRET Readout
      • Transcreener® ADO CD73 Assay Kit
      • Transcreener® AMP²/GMP² Phosphodiesterase Assay Kits
        • Transcreener® AMP2/GMP2 FP Assay
        • Transcreener® AMP2/GMP2 Assay Kit – TR-FRET Readout
      • Transcreener® cGAMP cGAS Assay Kits
        • Transcreener® cGAMP Assay Kit – FP Readout
        • Transcreener® cGAMP Assay Kit – TR-FRET Readout
      • Transcreener dAMP Exonuclease Assay Kit
      • Transcreener® EPIGEN SAH Methyltransferase Assay Kit
      • Transcreener® GDP GTPase Assay Kits
        • Transcreener® GDP Assay Kit – FP Readout
        • Transcreener® GDP Assay Kit – FI Readout
        • Transcreener® GDP Assay Kit – TR-FRET Readout
      • Transcreener® UDP² Glycosyltransferase Assay Kits
        • Transcreener® UDP2 Assay Kit – FP Readout
        • Transcreener® UDP2 Assay Kit – FI Readout
        • Transcreener® UDP2 Assay Kit – TR-FRET Readout
      • Transcreener® 2-5A OAS Assay Kit
    • AptaFluor® HTS Assay Kits
      • AptaFluor® SAH Methyltransferase Assay Kit
    • Enzyme Assay Systems
      • TREX1 Assay System
    • Recombinant Enzymes
      • Human cGAS Enzyme
      • Mouse cGAS Enzyme
      • Human DDX3 Enzyme
      • Human OAS1 Enzyme
      • Human TREX1 Enzyme
    • Assay Plates
    • Ordering Information
  • Services
    • Assay Development Services
    • Lead Discovery Services
    • CD38 Assay Services
    • GTPase Profiling Services
  • Assays by Target
    • Kinase Assays
      • ADK Assays – Application
      • AMPK Assays – Application
      • IKK-beta Assays – Application
      • IRAK4 Assays – Application
      • JAK1 Assays – Application
      • JAK3 Assays – Application
      • MAPK8 Assays – Application
      • PKR Assays – Application
      • RIPK1 Assays – Application
      • TBK1 Assays – Application
    • GTPase Assays
      • GAP Assays – Application
      • GEF Assays – Application
      • KRAS Assays – Application
      • HRAS Assays – Application
      • NRAS Assays – Application
      • RRAS Assays – Application
      • Rac1 Assays – Application
      • RhoA Assays – Application
      • RhoC Assays – Application
      • Cdc42 Assays – Application
      • Ran Assays – Application
    • Methyltransferase Assays
      • EZH2 Assays – Application
      • G9a Assays – Application
      • SET7/9 Assays – Application
      • SET8 Assays – Application
      • PRMT1 Assays – Application
      • PRMT3 Assays – Application
      • PRMT4 Assays – Application
    • STING Pathway Assays
      • cGAS Assay Kits
      • ENPP1 Assays – Application
      • TREX1 Assay System
      • IKK-beta Assays – Application
      • TBK1 Assays – Application
    • Nucleotidase Assays
      • CD38 Assay Services
      • CD39 Assays – Application
      • CD73 Activity Assay Kits
    • Helicase / ATPase Assays
      • DDX3 Assays – Application
      • NSP13 Assays – Application
      • P97 Assays – Application
    • Glycosyltransferase Assays
      • Toxin B Assays – Application
      • GALNT2 Assays – Application
      • GALNT3 Assays – Application
      • BGalT1 Assays – Application
    • Phosphodiesterase Assays
      • PDE3 Assays – Application
      • PDE4 Assays – Application
      • PDE5 Assays – Application
      • PDE7 Assays – Application
    • Ligase and Synthetase Assays
      • SUMO E1 Assays – Application
      • Acyl CoA Synthetase Assays – Application
      • S-Acetyl CoA Synthetase Assays – Application
    • Exonuclease Assays
      • TREX1 Assay System
    • OAS Assays
      • OAS1 Assay Kits
    • Other Enzyme Assays
      • NUDT5 Assays – Application
  • Resources
    • Technical Manuals
    • Transcreener® Assays – Instrument Compatibility
    • Application Notes
    • Posters and Presentations
    • Publications
    • Transcreener® FAQ’s
    • Guides
      • Residence Time Guide
      • Hit Prioritization Guide
      • Kinases in Innate Immunity
  • Company
    • President’s Message
    • International Distributors
    • Careers
    • Downloads
    • Contact Us
  • Blog
  • MY CART
    No products in cart.

The Multifaceted Roles of IRE1 Are Unfolding

by Bellbrook Labs / Wednesday, 09 March 2022 / Published in Emerging Targets, Innate Immunity
IRE1 Proteins in ER

IRE1 (Inositol-Requiring Enzyme 1) is one of three resident transmembrane endoplasmic reticulum (ER) proteins that sense and signal distress from improper protein synthesis and conformation. (While IRE1 alpha is almost universally expressed and IRE1 beta is confined to the gut, both will be conflated as IRE1 here.)  Secreted and transmembrane proteins are primarily produced in the lumen of the ER. When stressors, such as an accumulation of unfolded proteins, excessive protein synthesis, lipid overload, iron or calcium imbalance, hypoxia, oxidative stress, or viral infection occur, IRE1, along with PERK and ATF6, respond, via what is known as the Unfolded Protein Response (UPR), to restore ER homeostasis. Overall, this response adds “chaperone” molecules to help correct the folding of errant ER proteins, reduces the translational load in the ER via mRNA degradation in a process called regulated IRE1-dependent decay (RIDD), and destroys irredeemably unfolded protein via ERAD (ER-associated degradation). If the ER stress is short-lived, systems and cells return to nominal function. If the stress is chronic and severe, apoptotic or massively inflammatory situations evolve, triggering chronic disease.1

The IRE1 Mediated Adaptive UPR

The IRE1 protein spans the ER membrane, with its UPR sensing domain in the ER lumen and its catalytic region in the cytosol. IRE1’s catalytic region is composed of a serine/threonine kinase domain and an endoribonuclease domain. The Grp78 (aka BiP) protein is usually bound to luminal IRE1 under nominal conditions. When ER stress occurs, it dissociates from IRE1 and binds to unfolded proteins. This dissociation induces IRE1 oligomerization and autophosphorylation of the kinase domain. In turn, the endoribonuclease domain activates RIDD and performs novel splicing of XBP1 mRNA, yielding the transcription factor XBP1s.  XBP1s signals the nucleus to increase transcription of genes for protein folding chaperones, the ERAD system, and lipid biosynthesis. Depending on the particular cells and physiological conditions of ER stress, these upregulated genes affect secretion, lipid processing, glucose regulation, and inflammation.2

IRE1’s role in secreted protein production makes it vital to the health of a variety of highly secretory cells, including immune cells. The development, differentiation, activation, and cytokine expression of various immune cell types depend upon it. Upon viral or bacterial infection, the IRE1/XPB1 pathway contributes to the differentiation of CD8+ T cells into effector cells. By activating the c-Myc promoter, it also stimulates the expansion of natural killer cells. IRE1 and XPB1s are required for optimal production of pro-inflammatory cytokines in macrophages, with XPB1s essential for the differentiation of monocytes into macrophages. IRE1 is necessary for neutrophil infiltration, and active XBP1 is required for granule release. XPB1s are essential for B cells to differentiate into antibody-producing plasma cells and eosinophils to mature fully.3

The IRE1 Mediated Apoptotic UPR

However, under prolonged ER stress, activated IRE1 interacts with IKK via TRAF2, activating NFkB by degrading its inhibitor, IkBa.  IRE1 also binds TRAF2, activating ASK1 and, subsequently, JNK and p38 MAPK. JNK then upregulates pro-apoptotic genes, like BIM and PUMA, and upregulates AP-1. P38 MAPK promotes the transcription of CHOP axis genes, facilitating apoptosis. The now abundant NFkB and AP-1 stimulate the expression of potent pro-inflammatory cytokines (such as TNF alpha, IL-6, and IL beta), producing even more stress. Eventually, even XBP1s may begin to assist apoptotic CHOP induction. Oligomeric IRE1 can also induce the production of directly apoptotic Caspase 12.4

IRE1 in Disease

While the final disposition of IRE1 in chronic disease states is not yet known precisely, chronically stimulated and apoptotic IRE1 expression pathways are implicated in Alzheimer’s, Parkinson’s, and Huntington’s diseases,5 along with systemic lupus erythematosus. Just as in type 1 diabetes, loss of crucial cell subsets via apoptosis leads to severe impairments. IRE1 is involved with the autoimmune response to misfolded pro-insulin accumulation in Akita mice and misfolded HLA-B27 in human ankylosing spondylitis. IRE1/XBP1s induce high expression of Grp78 in rheumatoid arthritis, triggering a potent synovial T cell response. In inflammatory bowel disease, the IRE1/JNK axis upregulates NFkB, releasing damaging cytokines. The IRE1/RIDD axis degrades microRNAs that suppress type I and type IV collagen production in scleroderma, causing excess fibrosis.6 For researchers looking for IRE1 inhibitors the Transcreener Kinase Assay might be an option as it allows for the direct detection of ADP produced via IRE1 activity.

Slowly, the complex picture of IRE1’s necessity and liability is becoming more evident.

References

  1. So, Jae-Seon. (2018) Roles of Endoplasmic Reticulum Stress in Immune Responses. Molecules and Cells, 41(8), 705-716, Review. https://doi.org/10.14348/molcells.2018.0421.
  1. Hetz, C. et al. (2011) The Unfolded Protein Response: Integrating Stress Signals Through the Stress Sensor IRE1 alpha. Physiological Reviews, 91(4), 1219-1243, Review. https://doi.org/10.1152/physrev.00001.2011
  1. Coleman, O.I and Haller, D. (2019) ER Stress and the UPR in Shaping Intestinal Tissue Homeostasis and Immunity. Frontiers in Immunology, December 2019, Review. https://doi.org/10.3389/fimmu.2019.02825
  1. Ma, R-H et al. (2020) Natural Compounds Play Therapeutic Roles in Various Human Pathologies via Regulating Endoplasmic Reticulum Pathway. Medicine in Drug Discovery, vol. 8, December 2020, Review. https://doi.org/10.1016/j.medidd.2020.100065.
  1. Ni, H. et al. (2018) The Role of IRE1 Signaling in the Central Nervous System Diseases. Current Neuropharmacology, 16(9), 1340-1347, Review. https://pubmed.ncbi.nlm.nih.gov/29663887/
  1. Junjappa, R.P. et al. (2018) IRE1 alpha Implications in Endoplasmic Reticulum Stress-Mediated Development and Pathogenesis of Autoimmune Diseases. Frontiers in Immunology, Review. https://doi.org/10.3389/fimmu.2018.01289
Tagged under: Assay Development Services, drug discovery services, IRE1, Lead Discovery Services

What you can read next

Validation of a GEF Inhibitor in Murine Models of Major Osteolytic Diseases
tyrosine kinase inhibitor lung cancer
How a Tyrosine Kinase Inhibitor Is Giving Hope to Lung Cancer Patients
Glycosyltransferase Activity Assay and Lysosomal Storage Disorders
Using a Glycosyltransferase Activity Assay to Study Glycosylation and the Function of GTs

Categories

  • Company
  • Emerging Targets
  • Epigenetics
  • HTS Assays
  • Innate Immunity
  • Neurodegenerative Diseases
  • News
  • Products
  • Resources
  • Success Stories
  • Uncategorized

Recent Posts

  • SLAS 2023 Conference Exhibitor Announcement

    SLAS 2023 – HTS Assays and Discovery Services

    BellBrook Labs will exhibit and present posters...
  • BTK's Involved in Systemic lupus erythematosus

    The Challenging Search for BTK Inhibitors

    Bruton’s Tyrosine Kinase (BTK) is a 76kDa...
  • SARM1 Causes Axonal Death

    SARM1 Forefronts Research into Major Neurological Diseases

    SARM1 [Sterile alpha & toll/interleukin rec...
  • SIRT2 Affects Neurodegenerative Diseases

    SIRT2 Isoforms in Neurodegenerative Diseases & Cancer

    Silent information regulator type 2 (Sirtuin 2 ...
  • Patient With WRN Helicase Syndrome

    Can WRN Helicase Inhibitors Treat MSI-H Cancers?

    Over 100 years ago, Otto Werner first character...

Archives

BellBrook Labs
5500 Nobel Drive, Suite 230
Madison, Wisconsin 53711 USA
(608) 443-2400

info@bellbrooklabs.com

 Copyright © 2022 BellBrook Labs | All Rights Reserved | Privacy Policy | Terms of Use | FCOI | Sitemap

TOP