{"doi":"10.1007/978-1-0716-3377-9_13","title":"Single-Molecule FRET-Resolved Protein Dynamics – from Plasmid to Data in Six Steps","abstract":null,"journal":"Methods in Molecular Biology","year":2024,"id":596228,"datarank":0.20794415416798362,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"self_citation_contribution":0.20794415416798362,"citation_network_contribution":0.0,"self_endowment_contribution":0.20794415416798362,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"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":1526863,"name":"Jannick van Ossenbruggen","orcid":null,"position":1,"is_corresponding":false},{"id":207933,"name":"Sonja Schmid","orcid":null,"position":2,"is_corresponding":false},{"id":1526862,"name":"Benjamin Vermeer","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Single-Molecule FRET-Resolved Protein Dynamics – from Plasmid to Data in Six Steps","abstract":"Single-molecule Förster resonance energy transfer (smFRET) is a powerful technique for the detection of conformational dynamics of biomolecules. While many smFRET experiments are performed using dye-labeled DNA, here we describe a comprehensive protocol to resolve the conformational dynamics of a protein system - notably from plasmid to data. Using the example of the heat-shock protein Hsp90, we describe the protein production and threefold site-specific bioconjugation, the smFRET measurement using total internal reflection fluorescence microscopy (TIRFM), and raw data processing to reveal time-resolved protein dynamics. The described smFRET approach is readily transferrable to the study of many more all-protein systems and their conformational energy landscape.","is_dataset_classified":null,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"37824009","pmcid":null,"openalex_id":"https://openalex.org/W4387563295","authors":[],"funders":[],"total_grants":0,"fwci":2.0997,"citation_percentile":0.88467542,"influential_citations":0,"citation_trend":[{"year":2023,"count":1},{"year":2025,"count":2}],"oa_status":"green","license":"https://www.springernature.com/gp/researchers/text-and-data-mining","oa_locations":[{"url":"https://edepot.wur.nl/641764","host_type":"journal"},{"url":"https://edepot.wur.nl/641764","host_type":"repository"},{"url":"https://link.springer.com/content/pdf/10.1007/978-1-0716-3377-9_13","host_type":"publisher"},{"url":"https://research.wur.nl/en/publications/single-molecule-fret-resolved-protein-dynamics-from-plasmid-to-da","host_type":"journal"},{"url":"https://doi.org/10.1007/978-1-0716-3377-9_13","host_type":"book series"},{"url":"https://pubmed.ncbi.nlm.nih.gov/37824009","host_type":"repository"}],"fields_of_study":["Heat shock proteins research","ATP Synthase and ATPases Research","Advanced Fluorescence Microscopy Techniques","Fluorescence Resonance Energy Transfer","Molecular Conformation","Heat-Shock Proteins","Plasmids","DNA"],"mesh_terms":["DNA","Heat-Shock Proteins","Molecular Conformation","Plasmids","Fluorescence Resonance Energy Transfer"],"keywords":["Förster resonance energy transfer","Single-molecule FRET","Biophysics","Biomolecule","Total internal reflection fluorescence microscope","Protein dynamics","Dynamics (music)","Chemistry","Biological system","Protein structure","Fluorescence","Biology","Biochemistry","Physics","Protein purification","Bioconjugation","Surface Functionalization","Chaperone Protein Hsp90"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Affordable and clean energy"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-28T07:01:55.027884Z","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":[]}