{"doi":"10.1002/anie.202522420","title":"Investigation of Intercellular Trafficking of Global Glycosylphosphatidylinositol‐Anchored Proteins Using Cell Metabolic Engineering and Transwell Coculture","abstract":"Glycosylphosphatidylinositols (GPIs) are essential for GPI-anchored proteins (GPI-APs), as the loose association of GPIs with the cell membrane is thought to facilitate GPI-AP shedding and intercellular trafficking, which dictates cell communication. To explore this property, a novel method was developed based on metabolic engineering of GPI-APs in live cells and coculture of labelled donor cells along with recipient cells in a Transwell system for straight observation of intercellular GPI-AP transfer. Studies using this method have provided direct proof of intercellular GPI-AP trafficking and its dependence on cell types. Notably, there is a preference for GPI-AP transfer from cancer to normal cells. Lipid raft-mediated endocytosis and macropinocytosis play a critical role in GPI-AP trafficking and incorporation, with extracellular vesicles (EVs) involved as GPI-AP transporters. It is further revealed that depleting cholesterol in the donor cell can enhance GPI-AP shedding and trafficking. The method also allowed for in-depth studies to gain insights into the regulatory mechanisms of GPI-AP trafficking, such as its association with phospholipase A2 (PLA2), EV biogenesis, and membrane-adjacent Ras and Rho signaling pathways. The results are important not only for understanding GPI biology but also for its biomedical applications, for example, GPI-based cancer therapy.","journal":"Angewandte Chemie International Edition","year":2025,"id":537565,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9534,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":728369,"name":"Zachary T. Bennett","orcid":"0000-0001-9994-7939","position":1,"is_corresponding":false},{"id":433003,"name":"Jiatong Guo","orcid":null,"position":2,"is_corresponding":false},{"id":1310901,"name":"Palak Gupta","orcid":null,"position":3,"is_corresponding":false},{"id":407226,"name":"Zhongwu Guo","orcid":"0000-0001-5302-6456","position":4,"is_corresponding":false},{"id":1092229,"name":"Sayan Kundu","orcid":"0000-0003-0628-6698","position":0,"is_corresponding":true}],"reference_count":67,"raw_metadata":null,"created_at":"2026-07-19T02:52:12.997494Z","pmid":"41400405","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":[]}