{"doi":"10.1093/pnasnexus/pgac259","title":"Protein kinase R dependent phosphorylation of α-synuclein regulates its membrane binding and aggregation","abstract":"Aggregated α-synuclein (α-syn) accumulates in the neuronal Lewy body (LB) inclusions in Parkinson's disease (PD) and LB dementia. Yet, under nonpathological conditions, monomeric α-syn is hypothesized to exist in an equilibrium between disordered cytosolic- and partially α-helical lipid-bound states: a feature presumably important in synaptic vesicle release machinery. The exact underlying role of α-syn in these processes, and the mechanisms regulating membrane-binding of α-syn remains poorly understood. Herein we demonstrate that Protein kinase R (PKR) can phosphorylate α-syn at several Ser/Thr residues located in the membrane-binding region that is essential for α-syn's vesicle-interactions. α-Syn phosphorylated by PKR or α-syn isolated from PKR overexpressing cells, exhibit decreased binding to lipid membranes. Phosphorylation of Thr64 and Thr72 appears as the major contributor to this effect, as the phosphomimetic Thr64Glu/Thr72Glu-α-syn mutant displays reduced overall attachment to brain vesicles due to a decrease in vesicle-affinity of the last two thirds of α-syn's membrane binding region. This allows enhancement of the \"double-anchor\" vesicle-binding mechanism that tethers two vesicles and thus promote the clustering of presynaptic vesicles in vitro. Furthermore, phosphomimetic Thr64Glu/Thr72Glu-α-syn inhibits α-syn oligomerization and completely abolishes nucleation, elongation, and seeding of α-syn fibrillation in vitro and in cells, and prevents trans-synaptic spreading of aggregated α-syn pathology in organotypic hippocampal slice cultures. Overall, our findings demonstrate that normal and abnormal functions of α-syn, like membrane-binding, synaptic vesicle clustering and aggregation can be regulated by phosphorylation, e.g., via PKR. Mechanisms that could potentially be modulated for the benefit of patients suffering from α-syn aggregate-related diseases.","journal":"PNAS Nexus","year":2022,"id":263698,"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":13,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9528,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":642144,"name":"Hjalte Gram","orcid":"0000-0001-6516-5189","position":1,"is_corresponding":false},{"id":920740,"name":"Nanna Møller Jensen","orcid":"0000-0003-1177-5197","position":2,"is_corresponding":false},{"id":642148,"name":"Cristine Betzer","orcid":"0000-0001-5429-3548","position":3,"is_corresponding":false},{"id":920741,"name":"Yang Li","orcid":"0000-0001-7410-9111","position":4,"is_corresponding":false},{"id":921261,"name":"Lorrain Jin","orcid":null,"position":5,"is_corresponding":false},{"id":277410,"name":"Min Shi","orcid":"0000-0002-6901-2558","position":6,"is_corresponding":false},{"id":921262,"name":"Driss Boudeffa","orcid":null,"position":7,"is_corresponding":false},{"id":909421,"name":"Giuliana Fusco","orcid":"0000-0002-3644-9809","position":8,"is_corresponding":false},{"id":615301,"name":"Alfonso De Simone","orcid":"0000-0001-8789-9546","position":9,"is_corresponding":false},{"id":373683,"name":"Deniz Kirik","orcid":"0000-0002-1360-1265","position":10,"is_corresponding":false},{"id":225456,"name":"Hilal A. Lashuel","orcid":"0000-0001-7682-8320","position":11,"is_corresponding":false},{"id":707425,"name":"Jing Zhang","orcid":"0000-0002-9224-6299","position":12,"is_corresponding":false},{"id":625539,"name":"Poul Henning Jensen","orcid":"0000-0002-4439-9020","position":13,"is_corresponding":false},{"id":642147,"name":"Lasse Reimer","orcid":"0000-0002-6996-3185","position":0,"is_corresponding":true}],"reference_count":88,"raw_metadata":null,"created_at":"2026-07-19T00:26:33.413850Z","pmid":"36712380","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":[]}