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    • 20th Annual Phi Kappa Phi Student Research and Fine Arts Conference (2019)
    • 20th Annual PKP Student Research and Fine Arts Conference: Oral Symposia III
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    Unconventional Coupling of 5HT7 receptors to Gs heterotrimers

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    Authors
    Adams, Elizabeth
    Issue Date
    2019-02-13
    URI
    http://hdl.handle.net/10675.2/622093
    
    Metadata
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    Abstract
    GPCRs play a major role in cell signaling through their interactions with heterotrimeric G proteins. In conventional models of GPCR-G protein coupling, agonist binding promotes a conformational change within the receptor, which then associates with G proteins, facilitating the exchange of GDP for GTP. GTP-bound G proteins dissociate from the receptor and exert their effects on downstream signaling molecules. Previous studies suggest that serotonin 5HT7 receptors associate with Gs�heterotrimers prior to agonist binding, and that 5HT7-Gs�complexes dissociate after the G protein is activated. Here we study this unconventional mode of coupling using bioluminescence resonance energy transfer (BRET) between luciferase-tagged 5HT7 receptors and Gs�heterotrimers labeled with Venus. Our results confirm that 5HT7 receptors interact with inactive (GDP-bound) Gs�heterotrimers in the absence of an agonist, and that this interaction is stabilized by the inverse agonist methiothepin. Stimulation with the endogenous agonist serotonin (5HT) decreased BRET between 5HT7 receptors and Gs, indicating that the activation of the receptor leads to 5HT7-Gscomplex dissociation. Interestingly, Gs�activation was not required for complex dissociation. These results are consistent with the hypothesis that 5HT7 receptors couple to Gs�heterotrimers via an unconventional mechanism involving ligand-sensitive complexes of receptors and inactive Gs.
    Affiliation
    Department of Pharmacology and Toxicology
    Description
    Presentation given at the 20th Annual Phi Kappa Phi Student Research and Fine Arts Conference
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    Department of Chemistry and Physics: Student Research and Presentations
    Department of Pharmacology and Toxicology: Student Research and Presentations
    20th Annual PKP Student Research and Fine Arts Conference: Oral Symposia III

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