Research
Print page Print page
Switch language
The Capital Region of Denmark - a part of Copenhagen University Hospital
Published

A bioluminescence resonance energy transfer 2 (BRET2) assay for monitoring seven transmembrane receptor and insulin receptor crosstalk

Research output: Contribution to journalJournal articleResearchpeer-review

  1. A BRET assay for monitoring insulin receptor interactions and ligand pharmacology

    Research output: Contribution to journalJournal articleResearchpeer-review

  1. Functional brown adipose tissue and sympathetic activity after cold exposure in humans with type 1 narcolepsy

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. [Sar1, Ile4, Ile8]-angiotensin II Potentiates Insulin Receptor Signalling and Glycogen Synthesis in Hepatocytes

    Research output: Contribution to journalJournal articleResearchpeer-review

  3. Cerebrospinal fluid biomarkers of neurodegeneration are decreased or normal in narcolepsy

    Research output: Contribution to journalJournal articleResearchpeer-review

  4. Normal Morning MCH Levels and No Association with REM or NREM Sleep Parameters in Narcolepsy Type 1 and Type 2

    Research output: Contribution to journalJournal articleResearchpeer-review

  5. Monozygotic twins discordant for narcolepsy type 1 and multiple sclerosis

    Research output: Contribution to journalJournal articleResearchpeer-review

View graph of relations

The angiotensin AT1 receptor is a seven transmembrane (7TM) receptor, which mediates the regulation of blood pressure. Activation of angiotensin AT1 receptor may lead to impaired insulin signaling indicating crosstalk between angiotensin AT1 receptor and insulin receptor signaling pathways. To elucidate the molecular mechanisms behind this crosstalk, we applied the BRET2 technique to monitor the effect of angiotensin II on the interaction between Rluc8 tagged insulin receptor and GFP2 tagged insulin receptor substrates 1, 4, 5 (IRS1, IRS4, IRS5) and Src homology 2 domain-containing protein (Shc). We demonstrate that angiotensin II reduces the interaction between insulin receptor and IRS1 and IRS4, respectively, while the interaction with Shc is unaffected, and this effect is dependent on Gαq activation. Activation of other Gαq-coupled 7TM receptors led to a similar reduction in insulin receptor and IRS4 interactions whereas Gαs- and Gαi-coupled 7TM receptors had no effect. Furthermore, we used a panel of kinase inhibitors to show that angiotensin II engages different pathways when regulating insulin receptor interactions with IRS1 and IRS4. Angiotensin II inhibited the interaction between insulin receptor and IRS1 through activation of ERK1/2, while the interaction between insulin receptor and IRS4 was partially inhibited through protein kinase C dependent mechanisms. We conclude that the crosstalk between angiotensin AT1 receptor and insulin receptor signaling shows a high degree of specificity, and involves Gαq protein, and activation of distinct kinases. Thus, the BRET2 technique can be used as a platform for studying molecular mechanisms of crosstalk between insulin receptor and 7TM receptors.

Original languageEnglish
JournalJournal of Receptor and Signal Transduction Research
Volume37
Issue number6
Pages (from-to)590-599
Number of pages10
ISSN1079-9893
DOIs
Publication statusPublished - Dec 2017

    Research areas

  • Journal Article

ID: 52141834