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dc.contributor.authorFarooq, Maheen
dc.date.accessioned2019-02-13T20:07:19Z
dc.date.available2019-02-13T20:07:19Z
dc.date.issued2019-02-13
dc.identifier.urihttp://hdl.handle.net/10675.2/622119
dc.descriptionPresentation given at the 20th Annual Phi Kappa Phi Student Research and Fine Arts Conferenceen
dc.description.abstractGPCRs 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.
dc.subjectBRETen
dc.subjectG proteinsen
dc.subjectcouplingen
dc.titlePROFILING G PROTEINS USING BIOLUMINESCENCE RESONANCE ENERGY TRANSFERen
dc.typePoster Presentationen
dc.contributor.departmentDepartment of Chemistry and Physicsen
dc.contributor.departmentDepartment of Pharmacology & Toxicologyen
cr.funding.sourceAugusta University CURS Student Research Granten
dc.contributor.sponsorSpencer, Angelaen
dc.contributor.sponsorLambert, Nevinen
dc.contributor.sponsorOkashah, Najeahen
dc.contributor.affiliationAugusta Universityen
html.description.abstractGPCRs 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.


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