{"doi":"10.1038/s41598-021-87417-2","title":"New phosphosite-specific antibodies to unravel the role of GRK phosphorylation in dopamine D2 receptor regulation and signaling","abstract":"Abstract The dopamine D 2 receptor (D 2 R) is the target of drugs used to treat the symptoms of Parkinson’s disease and schizophrenia. The D 2 R is regulated through its interaction with and phosphorylation by G protein receptor kinases (GRKs) and interaction with arrestins. More recently, D 2 R arrestin-mediated signaling has been shown to have distinct physiological functions to those of G protein signalling. Relatively little is known regarding the patterns of D 2 R phosphorylation that might control these processes. We aimed to generate antibodies specific for intracellular D 2 R phosphorylation sites to facilitate the investigation of these mechanisms. We synthesised double phosphorylated peptides corresponding to regions within intracellular loop 3 of the hD 2 R and used them to raise phosphosite-specific antibodies to capture a broad screen of GRK-mediated phosphorylation. We identify an antibody specific to a GRK2/3 phosphorylation site in intracellular loop 3 of the D 2 R. We compared measurements of D 2 R phosphorylation with other measurements of D 2 R signalling to profile selected D 2 R agonists including previously described biased agonists. These studies demonstrate the utility of novel phosphosite-specific antibodies to investigate D 2 R regulation and signalling.","journal":"Scientific Reports","year":2021,"id":172444,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":25,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9595,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":463560,"name":"Alastair C. Keen","orcid":"0000-0002-0036-757X","position":1,"is_corresponding":false},{"id":709403,"name":"Hanka Mark","orcid":null,"position":2,"is_corresponding":false},{"id":708613,"name":"Pooja Dasgupta","orcid":"0000-0002-6369-122X","position":3,"is_corresponding":false},{"id":307532,"name":"Jonathan A. Javitch","orcid":"0000-0001-7395-2967","position":4,"is_corresponding":false},{"id":402418,"name":"Meritxell Canals","orcid":"0000-0002-7942-5006","position":5,"is_corresponding":false},{"id":443582,"name":"Stefan Schulz","orcid":"0000-0002-5997-8885","position":6,"is_corresponding":false},{"id":417632,"name":"J. Robert Lane","orcid":"0000-0002-7361-7875","position":7,"is_corresponding":false},{"id":708612,"name":"Anika Mann","orcid":"0000-0001-6500-4182","position":0,"is_corresponding":true}],"reference_count":59,"raw_metadata":null,"created_at":"2026-07-18T23:46:41.424716Z","pmid":"33859231","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":[]}