{"doi":"10.15252/msb.202311782","title":"Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification","abstract":"Phosphoprotein phosphatases (PPPs) regulate major signaling pathways, but the determinants of phosphatase specificity are poorly understood. This is because methods to investigate this at scale are lacking. Here, we develop a novel in vitro assay, MRBLE:Dephos, that allows multiplexing of dephosphorylation reactions to determine phosphatase preferences. Using MRBLE:Dephos, we establish amino acid preferences of the residues surrounding the dephosphorylation site for PP1 and PP2A-B55, which reveals common and unique preferences. To compare the MRBLE:Dephos results to cellular substrates, we focused on mitotic exit that requires extensive dephosphorylation by PP1 and PP2A-B55. We use specific inhibition of PP1 and PP2A-B55 in mitotic exit lysates coupled with phosphoproteomics to identify more than 2,000 regulated sites. Importantly, the sites dephosphorylated during mitotic exit reveal key signatures that are consistent with MRBLE:Dephos. Furthermore, integration of our phosphoproteomic data with mitotic interactomes of PP1 and PP2A-B55 provides insight into how binding of phosphatases to substrates shapes dephosphorylation. Collectively, we develop novel approaches to investigate protein phosphatases that provide insight into mitotic exit regulation.","journal":"Molecular Systems Biology","year":2023,"id":333511,"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":23,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.959,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":274544,"name":"Hieu Nguyen","orcid":"0000-0003-2153-9200","position":1,"is_corresponding":false},{"id":276408,"name":"Dimitriya H. Garvanska","orcid":null,"position":2,"is_corresponding":false},{"id":273954,"name":"Isha Nasa","orcid":"0000-0001-7699-795X","position":3,"is_corresponding":false},{"id":274541,"name":"Thomas Kruse","orcid":"0000-0002-2619-7388","position":4,"is_corresponding":false},{"id":491161,"name":"Yinnian Feng","orcid":null,"position":5,"is_corresponding":false},{"id":274546,"name":"Blanca López‐Méndez","orcid":"0000-0001-8541-4904","position":6,"is_corresponding":false},{"id":293148,"name":"Norman E. Davey","orcid":"0000-0001-6988-4850","position":7,"is_corresponding":false},{"id":273955,"name":"Arminja N. Kettenbach","orcid":"0000-0003-3979-4576","position":8,"is_corresponding":false},{"id":278296,"name":"Polly M. Fordyce","orcid":"0000-0002-9505-0638","position":9,"is_corresponding":false},{"id":274550,"name":"Jakob Nilsson","orcid":"0000-0003-4100-1125","position":10,"is_corresponding":false},{"id":274543,"name":"Jamin B. Hein","orcid":"0000-0002-2887-5082","position":0,"is_corresponding":true}],"reference_count":62,"raw_metadata":null,"created_at":"2026-07-19T01:09:39.719497Z","pmid":"37916966","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":[]}