{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T10:06:40Z","timestamp":1753870000706,"version":"3.41.2"},"reference-count":34,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T00:00:00Z","timestamp":1742169600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000015","name":"U.S. Department of Energy","doi-asserted-by":"publisher","award":["DE\u2010SC0012704"],"award-info":[{"award-number":["DE\u2010SC0012704"]}],"id":[{"id":"10.13039\/100000015","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R24GM145964","R35GM141933","S10_OD027000","S10_RR028976","S10_RR25528","UL1TR000454"],"award-info":[{"award-number":["R24GM145964","R35GM141933","S10_OD027000","S10_RR028976","S10_RR25528","UL1TR000454"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["R01EY021205"],"award-info":[{"award-number":["R01EY021205"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Glaucoma is a group of neurodegenerative diseases that together are the leading cause of irreversible blindness worldwide. Myocilin\u2010associated glaucoma is an inherited form of this disease, caused by intracellular aggregation of misfolded mutant myocilin. In vitro, the myocilin C\u2010terminal olfactomedin domain (OLF), the relevant domain for glaucoma pathogenesis, can be driven to form amyloid\u2010like fibrils under mild conditions. Here we characterize a species present during in vitro fibrillization. Purified OLF was subjected to fibrillization at concentrations required for downstream electron microscopy imaging and NMR spectroscopy. Additional biophysical techniques, including analytical ultracentrifugation and X\u2010ray crystallography, were employed to further characterize the multicomponent mixture. Negative stain transmission electron microscopy (TEM) shows a non\u2010native species reminiscent of known prefibrillar oligomers from other amyloid systems, NMR indicates a minor population of partially misfolded species is present in solution, and cryo\u2010EM imaging shows two\u2010dimensional protein arrays. The predominant soluble species remaining in solution after the fibril reaction is natively folded, as evidenced by X\u2010ray crystallography. In summary, after incubating OLF under fibrillization\u2010promoting conditions, there is a heterogeneous mixture consisting of soluble folded protein, mature amyloid\u2010like fibrils, and partially misfolded intermediate species that at present belie additional molecular detail. The characterization of OLF fibrillar species illustrates the challenges associated with developing a comprehensive understanding of the fibrillization process for large, non\u2010model amyloidogenic proteins.<\/jats:p>","DOI":"10.1002\/pro.70063","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T12:55:06Z","timestamp":1742216106000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Detection of non\u2010native species formed during fibrillization of the myocilin olfactomedin domain"],"prefix":"10.1002","volume":"34","author":[{"given":"Hailee F.","family":"Scelsi","sequence":"first","affiliation":[{"name":"School of Chemistry and Biochemistry Georgia Institute of Technology  Atlanta Georgia USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emily G. S.","family":"Close","sequence":"additional","affiliation":[{"name":"School of Chemistry and Biochemistry Georgia Institute of Technology  Atlanta Georgia USA"},{"name":"Pacific Northwest National Laboratory  Richland Washington USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dustin J. E.","family":"Huard","sequence":"additional","affiliation":[{"name":"School of Chemistry and Biochemistry Georgia Institute of Technology  Atlanta Georgia USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elijah","family":"Dunn","sequence":"additional","affiliation":[{"name":"School of Chemistry and Biochemistry Georgia Institute of Technology  Atlanta Georgia USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Neboj\u0161a","family":"Bogdanovi\u0107","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biophysics Florida State University  Tallahassee Florida USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sonali H. W.","family":"Mudiyanselage","sequence":"additional","affiliation":[{"name":"Biodesign Center for Personalized Diagnostics Arizona State University  Tempe Arizona USA"},{"name":"School of Molecular Sciences Arizona State University  Tempe Arizona USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arshay","family":"Grant","sequence":"additional","affiliation":[{"name":"School of Biological Sciences Georgia Institute of Technology  Atlanta Georgia USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Scott M.","family":"Stagg","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biophysics Florida State University  Tallahassee Florida USA"},{"name":"Department of Biological Sciences Florida State University  Tallahassee Florida USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ingeborg","family":"Schmidt\u2010Krey","sequence":"additional","affiliation":[{"name":"School of Biological Sciences Georgia Institute of Technology  Atlanta Georgia USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2493-5578","authenticated-orcid":false,"given":"Wade D.","family":"Van Horn","sequence":"additional","affiliation":[{"name":"Biodesign Center for Personalized Diagnostics Arizona State University  Tempe Arizona USA"},{"name":"School of Molecular Sciences Arizona State University  Tempe Arizona USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9345-3735","authenticated-orcid":false,"given":"Raquel L.","family":"Lieberman","sequence":"additional","affiliation":[{"name":"School of Chemistry and Biochemistry Georgia Institute of Technology  Atlanta Georgia USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,3,17]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/6.12.2091"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444912001308"},{"key":"e_1_2_8_4_1","doi-asserted-by":"crossref","unstructured":"BeplerT KelleyK NobleAJ BergerB.Topaz\u2010Denoise: general deep denoising models for cryoEM and cryoET.2020. bioRxiv 838920.","DOI":"10.1101\/838920"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0575-8"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-061516-045115"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00197809"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/ddu730"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.408906"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444910007493"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2012.01.006"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2018.01.002"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2013.12.002"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-48751-8"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2003.10.162"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1079469"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/0892-0354(90)90013-I"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/418291a"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444906045975"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/bm501521r"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2023.1120416"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2011.12.016"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(97)76066-X"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00249-023-01629-0"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep24526"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4169"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0502066102"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-44479-2"},{"key":"e_1_2_8_30_1","doi-asserted-by":"crossref","unstructured":"SaccuzzoEG RobangAS GaoY ChenB LiebermanRL ParavastuAK.Evidence for S331\u2010G\u2010S\u2010L within the amyloid core of myocilin olfactomedin domain fibrils based on low\u2010resolution 3D solid\u2010state NMR spectra.2024a. bioRxiv 2024.08.09.606901.","DOI":"10.1101\/2024.08.09.606901"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpc.2003.10.016"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10858-016-0060-y"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.exer.2016.07.009"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/ddx184"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.2353\/ajpath.2007.060806"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.70063","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.70063","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T04:55:30Z","timestamp":1745297730000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.70063"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,17]]},"references-count":34,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2025,4]]}},"alternative-id":["10.1002\/pro.70063"],"URL":"https:\/\/doi.org\/10.1002\/pro.70063","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"type":"print","value":"0961-8368"},{"type":"electronic","value":"1469-896X"}],"subject":[],"published":{"date-parts":[[2025,3,17]]},"assertion":[{"value":"2024-11-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-01-31","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-03-17","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70063"}}