{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T17:08:58Z","timestamp":1785431338670,"version":"3.56.0"},"reference-count":40,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T00:00:00Z","timestamp":1776211200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012190","name":"Ministry of Science and Higher Education of the Russian Federation","doi-asserted-by":"publisher","award":["075-15-2024-638"],"award-info":[{"award-number":["075-15-2024-638"]}],"id":[{"id":"10.13039\/501100012190","id-type":"DOI","asserted-by":"publisher"}]},{"award":["075-15-2024-638"],"award-info":[{"award-number":["075-15-2024-638"]}],"id":[{"id":"https:\/\/ror.org\/00ghqgy32","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Since the features of cross-seeding of alpha-synuclein forms may affect the sensitivity and specificity of the test systems, we developed a modified approach to obtain alpha-synuclein amyloid seeds with particle sizes from 20 to 50 nm prepared from either the wild-type protein (\u03b1-synWT) or its more fibrillation-prone form A53T (\u03b1-synA53T). These seeds had optimal properties for subsequent initiation of fibrillation. Our data showed that the elevated efficiency of alpha-synuclein A53T monomer transformation was hardly affected by the type of used seeds, whereas the addition of the seeds obtained from the alpha-synuclein mutant form to wild-type protein monomers had a significantly smaller effect than \u03b1-synWT seeds. Transmission electron microscopy data revealed that in the presence of \u03b1-synWT seeds the wild-type alpha-synuclein formed long and wide fibrils, while the addition of \u03b1-synA53T seeds led to the formation of long, but thin fibrils. Since the lag period of \u03b1-synA53T monomer fibrillation was significantly reduced compared to the wild-type protein, the replacing of \u03b1-synWT with \u03b1-synA53T in current assay systems designed to detect aberrant forms of \u03b1-synuclein in biological fluid samples (e.g., RT-QuIC) could substantially cut the time of analysis. In the future, a set of alpha-synuclein mutant forms could be used for the differential diagnosis of synucleinopathies caused by the different mutations of this protein.<\/jats:p>","DOI":"10.3390\/life16040675","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T10:30:23Z","timestamp":1776249023000},"page":"675","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Features of Cross-Seeding of Wild-Type Alpha-Synuclein and Its Mutant Form A53T Potentially Useful for the Development of Test Systems"],"prefix":"10.3390","volume":"16","author":[{"given":"Kseniya","family":"Barinova","sequence":"first","affiliation":[{"name":"Belozersky Research Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119991, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7549-607X","authenticated-orcid":false,"given":"Sofiya","family":"Kudryavtseva","sequence":"additional","affiliation":[{"name":"Belozersky Research Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119991, Russia"},{"name":"Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow 119991, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1030-0037","authenticated-orcid":false,"given":"Lidia","family":"Kurochkina","sequence":"additional","affiliation":[{"name":"Belozersky Research Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119991, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4877-1061","authenticated-orcid":false,"given":"Sergei","family":"Golyshev","sequence":"additional","affiliation":[{"name":"Belozersky Research Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119991, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7853-6222","authenticated-orcid":false,"given":"Nataliya","family":"Kolotyeva","sequence":"additional","affiliation":[{"name":"Russian Center of Neurology and Neurosciences, Moscow 125367, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sergei","family":"Illarioshkin","sequence":"additional","affiliation":[{"name":"Russian Center of Neurology and Neurosciences, Moscow 125367, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michail","family":"Piradov","sequence":"additional","affiliation":[{"name":"Russian Center of Neurology and Neurosciences, Moscow 125367, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladimir","family":"Muronetz","sequence":"additional","affiliation":[{"name":"Belozersky Research Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119991, Russia"},{"name":"Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow 119991, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2045","DOI":"10.1126\/science.276.5321.2045","article-title":"Mutation in the \u03b1-Synuclein Gene Identified in Families with Parkinson\u2019s Disease","volume":"276","author":"Polymeropoulos","year":"1997","journal-title":"Science"},{"key":"ref_2","first-page":"879","article-title":"Aggregation of Al-pha-Synuclein in Lewy Bodies of Sporadic Parkinson\u2019s Disease and Dementia with Lewy Bodies","volume":"152","author":"Baba","year":"1998","journal-title":"Am. J. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.cger.2019.08.002","article-title":"Parkinson Disease Epidemiology, Pathology, Genetics, and Pathophysiology","volume":"36","author":"Simon","year":"2020","journal-title":"Clin. Geriatr. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1186\/s13024-021-00501-z","article-title":"Neuropathology and molecular diagnosis of Synucleinopathies","volume":"16","author":"Koga","year":"2021","journal-title":"Mol. Neurodegener."},{"key":"ref_5","first-page":"1429","article-title":"Alpha-Synuclein Protofibrils in Cerebrospinal Fluid: A Potential Biomarker for Parkinson\u2019s Disease","volume":"10","author":"Giorgetti","year":"2020","journal-title":"J. Park. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2202","DOI":"10.1007\/s12035-020-02269-7","article-title":"Alpha-Synuclein Antibody Characterization: Why Semantics Matters","volume":"58","author":"Outeiro","year":"2021","journal-title":"Mol. Neurobiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1007\/s00401-017-1755-1","article-title":"Alpha-Synuclein Oligomers: A New Hope","volume":"134","author":"Roberts","year":"2017","journal-title":"Acta Neuropathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1038\/42166","article-title":"\u03b1-Synuclein in Lewy Bodies","volume":"388","author":"Spillantini","year":"1997","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"a024091","DOI":"10.1101\/cshperspect.a024091","article-title":"Cell Biology and Pathophysiology of \u03b1-Synuclein","volume":"8","author":"Sharma","year":"2018","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1517\/14728222.2015.1009448","article-title":"Targeting \u03b1-Synuclein Oligomers by Pro-tein-Fragment Complementation for Drug Discovery in Synucleinopathies","volume":"19","author":"Moussaud","year":"2015","journal-title":"Expert Opin. Ther. Targets"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Srinivasan, E., Chandrasekhar, G., Chandrasekar, P., Anbarasu, K., Vickram, A.S., Karunakaran, R., Rajasekaran, R., and Srikumar, P.S. (2021). Alpha-Synuclein Aggregation in Parkinson\u2019s Disease. Front. Med., 8.","DOI":"10.3389\/fmed.2021.736978"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1111\/nan.12298","article-title":"Review: Sporadic Parkinson\u2019s disease: Development and distribution of \u03b1-synuclein pathology","volume":"42","author":"Braak","year":"2016","journal-title":"Neuropathol. Appl. Neurobiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/S0140-6736(23)01478-2","article-title":"The Pathogenesis of Parkinson\u2019s Disease","volume":"403","author":"Morris","year":"2024","journal-title":"Lancet"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1002\/mds.27343","article-title":"Autonomic Dysfunction in Genetic Forms of Synucle-inopathies","volume":"33","author":"Chelban","year":"2018","journal-title":"Mov. Disord."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"112962","DOI":"10.1016\/j.celrep.2023.112962","article-title":"Mutant \u03b1-Synuclein Propagates via the Lymphatic System of the Brain in the Monomeric State","volume":"42","author":"Fujita","year":"2023","journal-title":"Cell Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/s40035-019-0164-x","article-title":"Efficient RT-QuIC Seeding Activity for \u03b1-Synuclein in Olfactory Mucosa Samples of Patients with Parkinson\u2019s Disease and Multiple System Atrophy","volume":"8","author":"Elia","year":"2019","journal-title":"Transl. Neurodegener."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e1944","DOI":"10.1212\/WNL.0000000000207147","article-title":"Misfolded \u03b1-Synuclein Assessment in the Skin and CSF by RT-QuIC in Isolated REM Sleep Behavior Disorder","volume":"100","author":"Iranzo","year":"2023","journal-title":"Neurology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1002\/acn3.338","article-title":"Alpha-Synuclein RT-QuIC in the CSF of Patients with Alpha-Synucleinopathies","volume":"3","author":"Fairfoul","year":"2016","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Orr\u00fa, C.D., Groveman, B.R., Hughson, A.G., Barrio, T., Isiofia, K., Race, B., Ferreira, N.C., Gambetti, P., Schneider, D.A., and Masujin, K. (2024). Sensitive detection of pathological seeds of \u03b1-synuclein, tau and prion protein on solid surfaces. PLoS Pathog., 20.","DOI":"10.1371\/journal.ppat.1012175"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/978-1-0716-1495-2_1","article-title":"\u03b1-Synuclein Seeding Assay Using RT-QuIC","volume":"2322","author":"Okuzumi","year":"2021","journal-title":"Methods Mol. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2120","DOI":"10.1002\/acn3.50897","article-title":"\u03b1-Synuclein RT-QuIC assay in cerebrospinal fluid of patients with dementia with Lewy bodies","volume":"6","author":"Bongianni","year":"2019","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e930","DOI":"10.1212\/WNL.0000000000012438","article-title":"Diagnostic Value of the CSF \u03b1-Synuclein Re-al-Time Quaking-Induced Conversion Assay at the Prodromal MCI Stage of Dementia With Lewy Bodies","volume":"97","author":"Rossi","year":"2021","journal-title":"Neurology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1002\/acn3.772","article-title":"Barcelona LRRK2 Study Group. \u03b1-Synuclein RT-QuIC in Cere-brospinal Fluid of LRRK2-Linked Parkinson\u2019s Disease","volume":"6","author":"Garrido","year":"2019","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1007\/s11481-019-09835-4","article-title":"Ul-trasensitive Detection of Aggregated \u03b1-Synuclein in Glial Cells, Human Cerebrospinal Fluid, and Brain Tissue Using the RT-QuIC Assay: New High-Throughput Neuroimmune Biomarker Assay for Parkinsonian Disorders","volume":"14","author":"Manne","year":"2019","journal-title":"J. Neuroimmune Pharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.ijbiomac.2016.12.011","article-title":"Dimerization of Tyr136Cys Alpha-Synuclein Prevents Amyloid Transformation of Wild Type Alpha-Synuclein","volume":"96","author":"Barinova","year":"2017","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1016\/j.bbrc.2022.07.015","article-title":"Effect of Bacteriophage-Encoded Chaperonins on Amyloid Transformation of \u03b1-Synuclein","volume":"622","author":"Leisi","year":"2022","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1038\/s41422-020-0299-4","article-title":"Cryo-EM Structure of Full-Length \u03b1-Synuclein Amyloid Fibril with Parkinson\u2019s Disease Familial A53T Mutation","volume":"30","author":"Sun","year":"2020","journal-title":"Cell Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1021\/acschemneuro.6b00287","article-title":"Distinct Mechanisms Determine \u03b1-Synuclein Fibril Morphology during Growth and Maturation","volume":"8","author":"Sidhu","year":"2017","journal-title":"ACS Chem. Neurosci."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Wilham, J.M., Orr\u00fa, C.D., Bessen, R.A., Atarashi, R., Sano, K., Race, B., Meade-White, K.D., Taubner, L.M., Timmes, A., and Caughey, B. (2010). Rapid end-point quantitation of prion seeding activity with sensitivity comparable to bioassays. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1001217"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/nm.2294","article-title":"Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion","volume":"17","author":"Atarashi","year":"2011","journal-title":"Nat. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s40478-018-0508-2","article-title":"Rapid and ultra-sensitive quantitation of disease-associated \u03b1-synuclein seeds in brain and cerebrospinal fluid by \u03b1Syn RT-QuIC","volume":"6","author":"Groveman","year":"2018","journal-title":"Acta Neuropathol. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1007\/s00401-017-1692-z","article-title":"Ultrasensitive and selective detection of 3-repeat tau seeding activity in Pick disease brain and cerebrospinal fluid","volume":"133","author":"Saijo","year":"2017","journal-title":"Acta Neuropathol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1007\/s00401-018-1947-3","article-title":"Seeding selectivity and ultrasensitive detection of tau aggregate conformers of Alzheimer disease","volume":"137","author":"Kraus","year":"2019","journal-title":"Acta Neuropathol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1007\/s00401-019-02080-2","article-title":"4-Repeat tau seeds and templating subtypes as brain and CSF biomarkers of frontotemporal lobar degeneration","volume":"139","author":"Saijo","year":"2020","journal-title":"Acta Neuropathol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1410","DOI":"10.1016\/j.bbadis.2019.02.013","article-title":"Sequence- and seed-structure-dependent polymorphic fibrils of alpha-synuclein","volume":"1865","author":"Tanaka","year":"2019","journal-title":"Biochim. Biophys. Acta (BBA)-Mol. Basis Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1074\/jbc.RA118.004471","article-title":"Parkinson\u2019s disease and multiple system atrophy have distinct \u03b1-synuclein seed characteristics","volume":"294","author":"Yamasaki","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1448","DOI":"10.1038\/s41591-023-02358-9","article-title":"Propagative \u03b1-synuclein seeds as serum biomarkers for synucleinopathies","volume":"29","author":"Okuzumi","year":"2023","journal-title":"Nat. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4906","DOI":"10.1093\/hmg\/ddx371","article-title":"Comparison of the in vivo induction and transmission of \u03b1-synuclein pathology by mutant \u03b1-synuclein fibril seeds in transgenic mice","volume":"26","author":"Rutherford","year":"2017","journal-title":"Hum. Mol. Genet"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Han, J.Y., Shin, C., and Choi, Y.P. (2021). Preclinical Detection of Alpha-Synuclein Seeding Activity in the Colon of a Transgenic Mouse Model of Synucleinopathy by RT-QuIC. Viruses, 13.","DOI":"10.3390\/v13050759"}],"container-title":["Life"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/4\/675\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T10:45:15Z","timestamp":1776249915000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/4\/675"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,15]]},"references-count":40,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["life16040675"],"URL":"https:\/\/doi.org\/10.3390\/life16040675","relation":{},"ISSN":["2075-1729"],"issn-type":[{"value":"2075-1729","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,15]]}}}