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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: Posters
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    PROFILING G PROTEINS USING BIOLUMINESCENCE RESONANCE ENERGY TRANSFER

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    Authors
    Farooq, Maheen
    Issue Date
    2019-02-13
    URI
    http://hdl.handle.net/10675.2/622119
    
    Metadata
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    Abstract
    GPCRs are receptors that act in signal transduction pathways via guanosine nucleotide-binding proteins (G proteins). Extracellular ligands act on GPCRs resulting in activation of one or more G protein subtypes (Gs, Gi/o, Gq/11 and G12/13) affecting the concentration of intracellular second messenger molecules ultimately altering cellular function. Cellular responses to external signals are typically studied indirectly by measuring concentration changes in second messengers. However, this approach can be problematic as many GPCRs can activate multiple G protein subtypes, and many second messenger pathways engage in crosstalk. To address this issue, we used Bioluminescence Resonance Energy Transfer (BRET) to directly measure coupling between 5-hydroxytryptamine (5-HT; serotonin) receptors and different G protein family subtypes. We co-transfected cells with plasmid DNA encoding the 5-HT2B or 5-HT4 receptors fused to the bioluminescent protein nanoluciferase (NLuc) as well as plasmid DNA containing G protein subtypes fused to the fluorescent protein Venus. In BRET assays, we found that mGsq couples to 5-HT2B and mGscouples with 5-HT4 in response to 5-HT activation. These results are consistent with the literature. Interestingly, initial studies suggest that activated 5-HT4 shows secondary coupling to mGsi highlighting the potential novel signaling pathways that can be elucidated using this technique.
    Affiliation
    Department of Chemistry and Physics
    Department of Pharmacology & Toxicology
    Description
    Presentation given at the 20th Annual Phi Kappa Phi Student Research and Fine Arts Conference
    Collections
    Department of Biological Sciences: Student Research and Presentations
    Department of Chemistry and Physics: Student Research and Presentations
    20th Annual PKP Student Research and Fine Arts Conference: Posters

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