{"doi":"10.1083/jcb.200308110","title":"Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles","abstract":"<jats:p>It has been thought that clathrin-mediated endocytosis is regulated by phosphorylation and dephosphorylation of many endocytic proteins, including amphiphysin I and dynamin I. Here, we show that Cdk5/p35-dependent cophosphorylation of amphiphysin I and dynamin I plays a critical role in such processes. Cdk5 inhibitors enhanced the electric stimulation–induced endocytosis in hippocampal neurons, and the endocytosis was also enhanced in the neurons of p35-deficient mice. Cdk5 phosphorylated the proline-rich domain of both amphiphysin I and dynamin I in vitro and in vivo. Cdk5-dependent phosphorylation of amphiphysin I inhibited the association with β-adaptin. Furthermore, the phosphorylation of dynamin I blocked its binding to amphiphysin I. The phosphorylation of each protein reduced the copolymerization into a ring formation in a cell-free system. Moreover, the phosphorylation of both proteins completely disrupted the copolymerization into a ring formation. Finally, phosphorylation of both proteins was undetectable in p35-deficient mice.</jats:p>","journal":"The Journal of Cell Biology","year":2003,"id":599942,"datarank":0.7954957362088615,"base_score":5.303304908059076,"endowment":5.303304908059076,"self_citation_contribution":0.7954957362088615,"citation_network_contribution":0.0,"self_endowment_contribution":0.7954957362088615,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":200,"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":1537797,"name":"Satoshi Sunada","orcid":null,"position":1,"is_corresponding":false},{"id":1537798,"name":"Yun-Fei Lu","orcid":null,"position":2,"is_corresponding":false},{"id":17618,"name":"Yoshiya Oda","orcid":null,"position":3,"is_corresponding":false},{"id":1526985,"name":"Masahiro Kinuta","orcid":null,"position":4,"is_corresponding":false},{"id":447096,"name":"Toshio Ohshima","orcid":"0000-0003-4931-7087","position":5,"is_corresponding":false},{"id":52526,"name":"Taro Saito","orcid":"0000-0003-4256-4173","position":6,"is_corresponding":false},{"id":1537799,"name":"Fan-Yan Wei","orcid":null,"position":7,"is_corresponding":false},{"id":385111,"name":"Masayuki Matsushita","orcid":"0000-0001-5373-3345","position":8,"is_corresponding":false},{"id":1537800,"name":"Sheng-Tian Li","orcid":null,"position":9,"is_corresponding":false},{"id":941561,"name":"Kimiko Tsutsui","orcid":null,"position":10,"is_corresponding":false},{"id":1537801,"name":"Shin-ichi Hisanaga","orcid":null,"position":11,"is_corresponding":false},{"id":134493,"name":"Katsuhiko Mikoshiba","orcid":null,"position":12,"is_corresponding":false},{"id":1526989,"name":"Kohji Takei","orcid":null,"position":13,"is_corresponding":false},{"id":1537802,"name":"Hideki Matsui","orcid":null,"position":14,"is_corresponding":false},{"id":650333,"name":"Kazuhito Tomizawa","orcid":"0000-0002-5663-2627","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles","abstract":"<jats:p>It has been thought that clathrin-mediated endocytosis is regulated by phosphorylation and dephosphorylation of many endocytic proteins, including amphiphysin I and dynamin I. Here, we show that Cdk5/p35-dependent cophosphorylation of amphiphysin I and dynamin I plays a critical role in such processes. Cdk5 inhibitors enhanced the electric stimulation–induced endocytosis in hippocampal neurons, and the endocytosis was also enhanced in the neurons of p35-deficient mice. Cdk5 phosphorylated the proline-rich domain of both amphiphysin I and dynamin I in vitro and in vivo. Cdk5-dependent phosphorylation of amphiphysin I inhibited the association with β-adaptin. Furthermore, the phosphorylation of dynamin I blocked its binding to amphiphysin I. The phosphorylation of each protein reduced the copolymerization into a ring formation in a cell-free system. Moreover, the phosphorylation of both proteins completely disrupted the copolymerization into a ring formation. Finally, phosphorylation of both proteins was undetectable in p35-deficient mice.</jats:p>","is_dataset_classified":null,"base_score":5.303304908059076,"endowment":5.303304908059076,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"14623869","pmcid":"PMC2173686","openalex_id":"https://openalex.org/W2098732177","authors":[],"funders":[],"total_grants":0,"fwci":6.3484,"citation_percentile":0.97596593,"influential_citations":0,"citation_trend":[{"year":2012,"count":13},{"year":2013,"count":5},{"year":2014,"count":8},{"year":2015,"count":4},{"year":2016,"count":2},{"year":2017,"count":5},{"year":2018,"count":3},{"year":2019,"count":2},{"year":2020,"count":4},{"year":2021,"count":4},{"year":2022,"count":1},{"year":2023,"count":6},{"year":2024,"count":3},{"year":2025,"count":3},{"year":2026,"count":4}],"oa_status":"bronze","license":null,"oa_locations":[{"url":"https://rupress.org/jcb/article-pdf/163/4/813/1312990/jcb1634813.pdf","host_type":"journal"},{"url":"https://rupress.org/jcb/article-pdf/163/4/813/1312990/jcb1634813.pdf","host_type":"publisher"},{"url":"https://rupress.org/jcb/article-pdf/163/4/813/1527523/jcb1634813.pdf","host_type":"publisher"},{"url":"https://doi.org/10.1083/jcb.200308110","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/14623869","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/2173686","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC2173686","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC2173686?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["Cellular transport and secretion","Lipid Membrane Structure and Behavior","Retinal Development and Disorders","Adaptor Protein Complex beta Subunits","Animals","Axonal Transport","Cells, Cultured","Clathrin","Cyclin-Dependent Kinase 5","Cyclin-Dependent Kinases","Dynamin I","Electric Stimulation","Endocytosis","Enzyme Inhibitors","Fetus","Hippocampus","Macromolecular Substances","Mice","Mice, Knockout","Nerve Tissue Proteins","Phosphorylation","Polymers","Protein Binding","Synaptic Vesicles"],"mesh_terms":["Animals","Axonal Transport","Cells, Cultured","Clathrin","Electric Stimulation","Endocytosis","Enzyme Inhibitors","Fetus","Hippocampus","Nerve Tissue Proteins","Phosphorylation","Polymers","Protein Binding","Synaptic Vesicles","Mice, Knockout","Cyclin-Dependent Kinases","Adaptor Protein Complex beta Subunits","Dynamin I","Macromolecular Substances","Cyclin-Dependent Kinase 5","Mice"],"keywords":["Amphiphysin","Dynamin","Endocytosis","Endocytic cycle","Phosphorylation","Cell biology","Clathrin","Chemistry","Cyclin-dependent kinase 5","Synaptic vesicle","Vesicle","Biology","Biochemistry","Cell","Protein kinase A","Membrane","Mitogen-activated protein kinase kinase"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-29T11:44:26.132391Z","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":[]}