{"doi":"10.1016/bs.mcb.2017.08.001","title":"Rapid kinetic BRET measurements to monitor G protein activation by GPCR and non-GPCR proteins","abstract":null,"journal":"Methods in Cell Biology","year":2017,"id":593778,"datarank":0.32958368660043297,"base_score":2.1972245773362196,"endowment":2.1972245773362196,"self_citation_contribution":0.32958368660043297,"citation_network_contribution":0.0,"self_endowment_contribution":0.32958368660043297,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1519796,"name":"Mikel Garcia-Marcos","orcid":null,"position":1,"is_corresponding":false},{"id":283571,"name":"Marcin Maziarz","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Rapid kinetic BRET measurements to monitor G protein activation by GPCR and non-GPCR proteins","abstract":"Heterotrimeric G proteins are central hubs of signal transduction whose activity is controlled by G protein-coupled receptors (GPCRs) as well as by a complex network of regulatory proteins. Recently, bioluminescence resonance energy transfer (BRET)-based assays have been used to monitor real-time activation of heterotrimeric G proteins in cells. Here we describe the use of a previously established BRET assay to monitor G protein activation upon GPCR stimulation and its adaptation to measure G protein activation by non-GPCR proteins, such as by cytoplasmic guanine nucleotide exchange factors (GEFs) like GIV/Girdin. The BRET assay monitors the release of free Gβγ from Gα-Gβγ heterotrimers as a readout of G protein activation, which is readily observable upon agonist stimulation of GPCRs. To control the signal input for non-GPCR activators, we describe the use of a chemically induced dimerization strategy to promote rapid membrane translocation of proteins containing the Gα-binding and -activating (GBA) motif found in some nonreceptor GEFs. The assay described here allows the kinetic measurement of G protein activation with subsecond temporal resolution and to compare the levels of activation induced by GPCR agonists vs those induced by the membrane recruitment of nonreceptor G protein signaling activators.","is_dataset_classified":null,"base_score":2.1972245773362196,"endowment":2.1972245773362196,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"28964333","pmcid":"PMC5654623","openalex_id":"https://openalex.org/W2755831226","authors":[],"funders":[{"funder_name":"NIGMS NIH HHS","grant_id":"R01 GM108733","title":null},{"funder_name":"NIGMS NIH HHS","grant_id":"R01 GM112631","title":null},{"funder_name":"National Institutes of Health","grant_id":"1R01GM112631-01","title":"Identification of chemical probes that specifically disrupt the GIV-Gi interface"},{"funder_name":"National Institutes of Health","grant_id":"5R01GM108733-03","title":"Alternative mechanisms of signaling via trimeric G proteins"}],"total_grants":4,"fwci":2.7683,"citation_percentile":0.90711217,"influential_citations":0,"citation_trend":[{"year":2017,"count":1},{"year":2018,"count":2},{"year":2019,"count":3},{"year":2020,"count":1},{"year":2021,"count":1}],"oa_status":"closed","license":"Elsevier TDM","oa_locations":[{"url":"https://api.elsevier.com/content/article/PII:S0091679X1730105X?httpAccept=text/xml","host_type":"publisher"},{"url":"https://api.elsevier.com/content/article/PII:S0091679X1730105X?httpAccept=text/plain","host_type":"publisher"},{"url":"https://doi.org/10.1016/bs.mcb.2017.08.001","host_type":"book series"},{"url":"https://pubmed.ncbi.nlm.nih.gov/28964333","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/5654623","host_type":"repository"},{"url":"https://europepmc.org/articles/pmc5654623?pdf=render","host_type":""},{"url":"https://dx.doi.org/10.1016/bs.mcb.2017.08.001","host_type":""}],"fields_of_study":["bioluminescence and chemiluminescence research","Receptor Mechanisms and Signaling","Retinal Development and Disorders","0301 basic medicine","0303 health sciences","03 medical and health sciences","Amino Acid Motifs","Bioluminescence Resonance Energy Transfer Techniques","Cell Membrane","GTP-Binding Proteins","HEK293 Cells","Heterotrimeric GTP-Binding Proteins","Humans","Kinetics","Microfilament Proteins","Protein Multimerization","Signal Transduction","Time Factors","Vesicular Transport Proteins"],"mesh_terms":["Cell Membrane","Humans","Kinetics","Microfilament Proteins","Time Factors","Signal Transduction","GTP-Binding Proteins","Amino Acid Motifs","Heterotrimeric GTP-Binding Proteins","Vesicular Transport Proteins","Protein Multimerization","HEK293 Cells","Bioluminescence Resonance Energy Transfer Techniques"],"keywords":["Heterotrimeric G protein","G protein-coupled receptor","G protein","Chemistry","Cell biology","Signal transduction","Biophysics","Biochemistry","Biology","Gpcr","Protein–protein Interaction","Gef","Bioluminescence Resonance Energy Transfer","Giv/girdin","Bioluminescence Resonance Energy Transfer Techniques","Time Factors","Amino Acid Motifs","Cell Membrane","Microfilament Proteins","Vesicular Transport Proteins","Heterotrimeric GTP-Binding Proteins","Kinetics","HEK293 Cells","GTP-Binding Proteins","Humans","Protein Multimerization"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Affordable and clean energy"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-27T12:12:15.611349Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}