{"doi":"10.1074/jbc.m114.567834","title":"A Novel Regulatory Locus of Phosphorylation in the C Terminus of the Potassium Chloride Cotransporter KCC2 That Interferes with N-Ethylmaleimide or Staurosporine-mediated Activation*♦","abstract":null,"journal":"Journal of Biological Chemistry","year":2014,"id":686853,"datarank":0.5742962094733643,"base_score":3.828641396489095,"endowment":3.828641396489095,"self_citation_contribution":0.5742962094733643,"citation_network_contribution":0.0,"self_endowment_contribution":0.5742962094733643,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":45,"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":1791412,"name":"Anna-Maria Hartmann","orcid":null,"position":1,"is_corresponding":false},{"id":1794376,"name":"Timo Beyer","orcid":null,"position":2,"is_corresponding":false},{"id":1794377,"name":"Anne Ripperger","orcid":null,"position":3,"is_corresponding":false},{"id":1791414,"name":"Hans Gerd Nothwang","orcid":null,"position":4,"is_corresponding":false},{"id":1794375,"name":"Maren Weber","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"A Novel Regulatory Locus of Phosphorylation in the C Terminus of the Potassium Chloride Cotransporter KCC2 That Interferes with N-Ethylmaleimide or Staurosporine-mediated Activation*♦","abstract":"The neuron-specific cation chloride cotransporter KCC2 plays a crucial role in hyperpolarizing synaptic inhibition. Transporter dysfunction is associated with various neurological disorders, raising interest in regulatory mechanisms. Phosphorylation has been identified as a key regulatory process. Here, we retrieved experimentally observed phosphorylation sites of KCC2 from public databases and report on the systematic analysis of six phosphorylated serines, Ser(25), Ser(26), Ser(937), Ser(1022), Ser(1025), and Ser(1026). Alanine or aspartate substitutions of these residues were analyzed in HEK-293 cells. All mutants were expressed in a pattern similar to wild-type KCC2 (KCC2(WT)). Tl(+) flux measurements demonstrated unchanged transport activity for Ser(25), Ser(26), Ser(1022), Ser(1025), and Ser(1026) mutants. In contrast, KCC2(S937D), mimicking phosphorylation, resulted in a significant up-regulation of transport activity. Aspartate substitution of Thr(934), a neighboring putative phosphorylation site, resulted in a comparable increase in KCC2 transport activity. Both KCC2(T934D) and KCC2(S937D) mutants were inhibited by the kinase inhibitor staurosporine and by N-ethylmaleimide, whereas KCC2(WT), KCC2(T934A), and KCC2(S937A) were activated. The inverse staurosporine effect on aspartate versus alanine substitutions reveals a cross-talk between different phosphorylation sites of KCC2. Immunoblot and cell surface labeling experiments detected no alterations in total abundance or surface expression of KCC2(T934D) and KCC2(S937D) compared with KCC2(WT). These data reveal kinetic regulation of transport activity by these residues. In summary, our data identify a novel key regulatory phosphorylation site of KCC2 and a functional interaction between different conformation-changing post-translational modifications. The action of pharmacological agents aimed to modulate KCC2 activity for therapeutic benefit might therefore be highly context-specific.","is_dataset_classified":null,"base_score":3.828641396489095,"endowment":3.828641396489095,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"24849604","pmcid":"PMC4081912","openalex_id":"https://openalex.org/W1992851684","authors":[],"funders":[],"total_grants":0,"fwci":1.5178,"citation_percentile":0.78651917,"influential_citations":0,"citation_trend":[{"year":2014,"count":1},{"year":2015,"count":5},{"year":2017,"count":6},{"year":2018,"count":3},{"year":2019,"count":5},{"year":2020,"count":7},{"year":2021,"count":4},{"year":2022,"count":4},{"year":2023,"count":6},{"year":2024,"count":1},{"year":2025,"count":2},{"year":2026,"count":1}],"oa_status":"hybrid","license":"cc-by","oa_locations":[{"url":"http://www.jbc.org/article/S0021925820404600/pdf","host_type":"journal"},{"url":"http://www.jbc.org/article/S0021925820404600/pdf","host_type":"publisher"},{"url":"https://api.elsevier.com/content/article/PII:S0021925820404600?httpAccept=text/xml","host_type":"publisher"},{"url":"https://api.elsevier.com/content/article/PII:S0021925820404600?httpAccept=text/plain","host_type":"publisher"},{"url":"https://syndication.highwire.org/content/doi/10.1074/jbc.M114.567834","host_type":"publisher"},{"url":"https://doi.org/10.1074/jbc.m114.567834","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/24849604","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/4081912","host_type":"repository"},{"url":"http://oops.uni-oldenburg.de/2350/","host_type":"repository"},{"url":"https://doi.org/10.1074/jbc.M114.567834","host_type":"repository"}],"fields_of_study":["Neuroscience and Neuropharmacology Research","Ion channel regulation and function","Molecular Sensors and Ion Detection"],"mesh_terms":["K Cl- Cotransporters","Amino Acid Sequence","Animals","Binding Sites","Ethylmaleimide","Gene Expression Regulation","Humans","Molecular Sequence Data","Mutation","Phosphoproteins","Phosphorylation","Phylogeny","Staurosporine","Amino Acid Substitution","Protein Transport","Symporters","Databases, Protein","Mice","Rats","Data Mining","HEK293 Cells"],"keywords":["Phosphorylation","Staurosporine","Cotransporter","Symporter","Mutant","Biochemistry","Chemistry","Alanine","Cell biology","Protein kinase A","Protein phosphorylation","Biology","Transporter","Gene","Amino acid","Sodium","Mutation","Evolution","Neurophysiology","Protein conformation","Cell culture","Post-translational modification","Chloride Transport","Cation Chloride Cotransporter"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"refseq"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-18T20:40:10.219491Z","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":[]}