{"doi":"10.1038/s42003-020-0840-5","title":"Transmembrane protein rotaxanes reveal kinetic traps in the refolding of translocated substrates","abstract":"Understanding protein folding under conditions similar to those found in vivo remains challenging. Folding occurs mainly vectorially as a polypeptide emerges from the ribosome or from a membrane translocon. Protein folding during membrane translocation is particularly difficult to study. Here, we describe a single-molecule method to characterize the folded state of individual proteins after membrane translocation, by monitoring the ionic current passing through the pore. We tag both N and C termini of a model protein, thioredoxin, with biotinylated oligonucleotides. Under an electric potential, one of the oligonucleotides is pulled through a α-hemolysin nanopore driving the unfolding and translocation of the protein. We trap the protein in the nanopore as a rotaxane-like complex using streptavidin stoppers. The protein is subjected to cycles of unfolding-translocation-refolding switching the voltage polarity. We find that the refolding pathway after translocation is slower than in bulk solution due to the existence of kinetic traps.","journal":"Communications Biology","year":2020,"id":81036,"datarank":0.40620753016533157,"base_score":2.70805020110221,"endowment":2.70805020110221,"self_citation_contribution":0.40620753016533157,"citation_network_contribution":0.0,"self_endowment_contribution":0.40620753016533157,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":14,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9615,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":421064,"name":"Pablo Martin-Baniandres","orcid":null,"position":1,"is_corresponding":false},{"id":419723,"name":"Michael J. Booth","orcid":"0000-0002-4224-798X","position":2,"is_corresponding":false},{"id":419724,"name":"Gianluca Veggiani","orcid":"0000-0002-4064-0112","position":3,"is_corresponding":false},{"id":419725,"name":"Mark Howarth","orcid":"0000-0001-8870-7147","position":4,"is_corresponding":false},{"id":380237,"name":"Hagan Bayley","orcid":"0000-0003-2499-6116","position":5,"is_corresponding":false},{"id":380238,"name":"David Rodríguez‐Larrea","orcid":"0000-0002-5580-7413","position":6,"is_corresponding":false},{"id":421063,"name":"Jianfei Feng","orcid":null,"position":0,"is_corresponding":true}],"reference_count":58,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T21:52:34.401799Z","pmid":"32246060","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":[]}