{"doi":"10.3233/jpd-240296","title":"Structurally targeted mutagenesis identifies key residues supporting α-synuclein misfolding in multiple system atrophy","abstract":"Background Multiple system atrophy (MSA) and Parkinson's disease (PD) are caused by misfolded α-synuclein spreading throughout the central nervous system. While familial PD is linked to several α-synuclein mutations, no mutations are associated with MSA. We previously showed that the familial PD mutation E46K inhibits replication of MSA prions both in vitro and in vivo , providing key evidence to support the hypothesis that α-synuclein adopts unique strains in patients. Objective Here we sought to further interrogate α-synuclein misfolding to identify the structural determinants that contribute to MSA strain biology. Methods We engineered a panel of cell lines harbouring both PD-linked and novel mutations designed to identify key residues that facilitate α-synuclein misfolding in MSA. We also used Maestro in silico analyses to predict the effect of each mutation on α-synuclein misfolding into one of the reported MSA cryo-electron microscopy conformations. Results In many cases, our modelling accurately identified mutations that facilitated or inhibited MSA replication. However, Maestro was occasionally unable to predict the effect of a mutation, demonstrating the challenge of using computational tools to investigate intrinsically disordered proteins. Finally, we used our cellular models to determine the mechanism underlying the E46K-driven inhibition of MSA replication, finding that the E46/K80 salt bridge is necessary to support α-synuclein misfolding. Conclusions Our studies used a structure-based approach to investigate α-synuclein misfolding, resulting in the creation of a powerful panel of cell lines that can be used to interrogate MSA strain biology.","journal":"Journal of Parkinson s Disease","year":2024,"id":456841,"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":6,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9542,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":914182,"name":"Sara A. M. Holec","orcid":"0000-0003-0800-1115","position":1,"is_corresponding":false},{"id":1116889,"name":"Chimere Ezeiruaku","orcid":null,"position":2,"is_corresponding":false},{"id":1080700,"name":"Matthew P. Frost","orcid":"0000-0002-9973-2572","position":3,"is_corresponding":false},{"id":409708,"name":"Christine Brown","orcid":"0000-0001-8771-3713","position":4,"is_corresponding":false},{"id":1052030,"name":"Samantha L Liu","orcid":"0009-0007-3692-0879","position":5,"is_corresponding":false},{"id":403981,"name":"Steven H. Olson","orcid":"0000-0001-9615-9355","position":6,"is_corresponding":false},{"id":471322,"name":"Amanda L. Woerman","orcid":"0000-0003-4029-1968","position":7,"is_corresponding":false},{"id":446348,"name":"Patrícia Reis","orcid":null,"position":0,"is_corresponding":true}],"reference_count":51,"raw_metadata":null,"created_at":"2026-07-19T02:03:32.516678Z","pmid":"39957201","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":[]}