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    The Study of 5-HT1D and 5-HT1F Receptor Interactions with G Proteins via BRET Analysis

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    Trang, A. (Thesis Final).pdf
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    Authors
    Trang, Amy
    Issue Date
    2019-05
    URI
    http://hdl.handle.net/10675.2/622413
    
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    Abstract
    G protein-coupled receptors (GPCRs) are receptors involved in signal transduction, a process for converting extracellular signals into internal messages to elicit a cellular response. Signal transduction pathways involve activating various G protein subtypes (Gs, Gi/o, Gq/11 and G12/13) which typically lead to second messenger production. Traditionally, second messenger concentration assays are used to identify GPCR coupling with G protein(s), but they are not efficient in profiling GPCRs since they compare the concentrations from different downstream signals. Instead, novel tools, such as Bioluminescence Resonance Energy Transfer (BRET) and mini G (mG) proteins, can be used to profile GPCRs. BRET is a technique that provides quantitative data when protein-protein interaction occurs and requires the proteins of interest to be fused with either a bioluminescent protein or fluorescent protein. In this study, we used mG proteins representing each G protein subtype to identify 5-hydroxytryptamine (5-HT; serotonin) receptor coupling upon serotonin stimulation. Through BRET assays, we determined that both the 5-HT1D and 5-HT1F receptors couple primarily with the mGsiand mGo classes of mG proteins. This supports previous studies that these receptors couple to Gi/o proteins and suggests that the use of mG proteins in BRET assays is an effective tool for GPCR profiling.
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    Department of Biological Sciences
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    Department of Biological Sciences: Student Research and Presentations
    Honors Program Theses

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