{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/89216"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/89216","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Solid-state NMR studies of α-synuclein fibril structure","abstract":"Lewy bodies are intracytoplasmic aggregates that are the major pathological feature of Parkinson’s disease (PD). Their primary protein component, α-synuclein (α- syn), adopts an amyloidogenic fibrillar form in Lewy bodies. In recent years research has slowly started to unravel clear pathological links between this fibrillar protein and PD disease progression. While great progress is being made, it has become evident that the structures of different α-syn fibril forms are closely linked to their pathogenicity, and the lack of structural details have hindered our understanding of PD. Here, I present the first 3D structure of α-syn fibrils determined through solid- state NMR spectroscopy experiments validated with X-ray fiber diffraction and electron microscopy mass per-length measurements. The fibril form presented here is shown to be pathologically active in neuronal cell culture. Further, the structure is validated by comparison of the structure to three of the early-onset PD mutants, A30P, E46K, and A53T, and show that the changes in the SSNMR spectra induced by the mutation are consistent with the structural features present in the fibril, most notably the E46K mutant disrupting a stabilizing salt-bridge within the fibril core. Additionally, I show that non- native cysteine mutants of T33C, V48C, T59C, A85C, and Q99C further validate the structure. The results and studies presented here establish the first structure of α-syn fibrils and provide the experimental and computational basis for greatly increasing our understanding of PD, amyloid fibril structure, and advancing the techniques available for structural studies via SSNMR spectroscopy.","abstract_html":"Lewy bodies are intracytoplasmic aggregates that are the major pathological feature of Parkinson’s disease (PD). Their primary protein component, α-synuclein (α- syn), adopts an amyloidogenic fibrillar form in Lewy bodies. In recent years research has slowly started to unravel clear pathological links between this fibrillar protein and PD disease progression. While great progress is being made, it has become evident that the structures of different α-syn fibril forms are closely linked to their pathogenicity, and the lack of structural details have hindered our understanding of PD. Here, I present the first 3D structure of α-syn fibrils determined through solid- state NMR spectroscopy experiments validated with X-ray fiber diffraction and electron microscopy mass per-length measurements. The fibril form presented here is shown to be pathologically active in neuronal cell culture. Further, the structure is validated by comparison of the structure to three of the early-onset PD mutants, A30P, E46K, and A53T, and show that the changes in the SSNMR spectra induced by the mutation are consistent with the structural features present in the fibril, most notably the E46K mutant disrupting a stabilizing salt-bridge within the fibril core. Additionally, I show that non- native cysteine mutants of T33C, V48C, T59C, A85C, and Q99C further validate the structure. The results and studies presented here establish the first structure of α-syn fibrils and provide the experimental and computational basis for greatly increasing our understanding of PD, amyloid fibril structure, and advancing the techniques available for structural studies via SSNMR spectroscopy.","abstract_has_math":false,"creators":["Tuttle, Marcus Dale"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Chemistry","degree_department":null,"school":null,"contributors":["Rienstra, Chad M","Murphy, Catherine J","Bailey, Ryan C","Hergenrother, Paul J"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2016,"date_issued":"2016-03-02T21:06:50Z","date_published":"2016-03-02T21:06:50Z","updated_at":"2026-07-22T22:26:32Z","subjects":["solid state nuclear magnetic resonance (SSNMR)","α-Synuclein","Parkinson's Disease","Nuclear magnetic resonance (NMR)","Structural Biology"],"languages":["en"],"rights":["Copyright 2015 Marcus Dale Tuttle"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/89216","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Rienstra, Chad M","Murphy, Catherine J","Bailey, Ryan C","Hergenrother, Paul J"]},{"key":"dc:creator","label":"Author","values":["Tuttle, Marcus Dale"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2016-03-02T21:06:50Z","2018-03-03T10:15:22Z","2015-12-01","2015-12"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["solid state nuclear magnetic resonance (SSNMR)","α-Synuclein","Parkinson's Disease","Nuclear magnetic resonance (NMR)","Structural Biology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2015 Marcus Dale Tuttle"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/89216"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Lewy bodies are intracytoplasmic aggregates that are the major pathological feature of Parkinson’s disease (PD). Their primary protein component, α-synuclein (α- syn), adopts an amyloidogenic fibrillar form in Lewy bodies. In recent years research has slowly started to unravel clear pathological links between this fibrillar protein and PD disease progression. While great progress is being made, it has become evident that the structures of different α-syn fibril forms are closely linked to their pathogenicity, and the lack of structural details have hindered our understanding of PD. Here, I present the first 3D structure of α-syn fibrils determined through solid- state NMR spectroscopy experiments validated with X-ray fiber diffraction and electron microscopy mass per-length measurements. The fibril form presented here is shown to be pathologically active in neuronal cell culture. Further, the structure is validated by comparison of the structure to three of the early-onset PD mutants, A30P, E46K, and A53T, and show that the changes in the SSNMR spectra induced by the mutation are consistent with the structural features present in the fibril, most notably the E46K mutant disrupting a stabilizing salt-bridge within the fibril core. Additionally, I show that non- native cysteine mutants of T33C, V48C, T59C, A85C, and Q99C further validate the structure. The results and studies presented here establish the first structure of α-syn fibrils and provide the experimental and computational basis for greatly increasing our understanding of PD, amyloid fibril structure, and advancing the techniques available for structural studies via SSNMR spectroscopy.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2017-12-01","The student, Marcus Tuttle, accepted the attached license on 2015-11-30 at 16:03.","The student, Marcus Tuttle, submitted this Dissertation for approval on 2015-11-30 at 16:23.","This Dissertation was approved for publication on 2015-12-01 at 12:09.","DSpace SAF Submission Ingestion Package generated from Vireo submission #8877 on 2016-03-02 at 14:13:31","Made available in DSpace on 2016-03-02T21:06:50Z (GMT). No. of bitstreams: 20 TUTTLE-DISSERTATION-2015.pdf: 25957302 bytes, checksum: e0ff2c528ec0f9585177d1b1ff5766e6 (MD5) Chapter_0a_Title_Page_SPACING.docx: 46703 bytes, checksum: 626a773119041caf0972cd994bb7b820 (MD5) Chapter_0b_Abstract_DONE.docx: 115185 bytes, checksum: 28042ff974493c824b35b8d1c027e05d (MD5) Chapter_0c_Acknowledgements_DONE.docx: 119424 bytes, checksum: 5f4b49133be9e5db06b20f28c354d30d (MD5) Chapter_0d_TableofContents_NEEDS_PAGENUMS.docx: 81655 bytes, checksum: 588b3ee9e1cc86614cf220f84c93f66c (MD5) Chapter_1_DONE.docx: 1240527 bytes, checksum: 500c637f4063b494a254b5cbc6b2e173 (MD5) Chapter_2_DONE.docx: 943435 bytes, checksum: a8b7dc731f90a4b090a07e2b3a655eb9 (MD5) Chapter_3_DONE.docx: 1840648 bytes, checksum: 029f682f63995ffbeeea2f070f4a950e (MD5) Chapter_4_DONE.docx: 867234 bytes, checksum: 492e11369a275b830feee8abf0fb1dcf (MD5) Chapter_5_DONE.docx: 10890464 bytes, checksum: 8baa4cee07aa6c6567a1f357e5d00361 (MD5) Chapter_6_DONE_18NOV15.docx: 1093414 bytes, checksum: 62d6ce9cfb681a7daf48c0e34238110c (MD5) Chapter_7_DONE.docx: 303105 bytes, checksum: 57a825c3a8302d5112b847056102a112 (MD5) Chapter_8_DONE.docx: 139574 bytes, checksum: 9ff6fff4015dc3c389f461fdf9c47cd2 (MD5) Chapter3_MiMB_License.pdf: 287085 bytes, checksum: 5d3cca17ca676720df9faa4d7f080746 (MD5) Chapter_1_Fig1.pdf: 574926 bytes, checksum: b39a6f39b6a4da71962ba9f3fb6174ec (MD5) Chapter_1_Fig2.pdf: 46094 bytes, checksum: 73bcec5a60f1aa95e345344264496cd4 (MD5) Chapter_3_Fig_9.pdf: 136509 bytes, checksum: 7f3c5bd517df90d85e5221baaa5cb85d (MD5) Chapter_6_Fig1.pdf: 668338 bytes, checksum: b2a33700b3e9951fdd41c29053bb1e55 (MD5) Glycerol_Labeling_Permission.pdf: 196227 bytes, checksum: 969a8f5f2e89a2d26e19ab5e5cd800ba (MD5) LICENSE.txt: 4210 bytes, checksum: 30579cafc7a0159016efb81c25b31cc5 (MD5) Previous issue date: 2015-12-01","Embargo set by: Seth Robbins for item 91419 Lift date: 2018-03-02T21:07:27Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Limited Restriction Lifted for Item 91419 on 2018-03-03T10:15:22Z."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Solid-state NMR studies of α-synuclein fibril structure"]}]}],"canonical_facts":{"dc:contributor":["Rienstra, Chad M","Murphy, Catherine J","Bailey, Ryan C","Hergenrother, Paul J"],"dc:creator":["Tuttle, Marcus Dale"],"dc:date":["2016-03-02T21:06:50Z","2018-03-03T10:15:22Z","2015-12-01","2015-12"],"dc:description":["Lewy bodies are intracytoplasmic aggregates that are the major pathological feature of Parkinson’s disease (PD). Their primary protein component, α-synuclein (α- syn), adopts an amyloidogenic fibrillar form in Lewy bodies. In recent years research has slowly started to unravel clear pathological links between this fibrillar protein and PD disease progression. While great progress is being made, it has become evident that the structures of different α-syn fibril forms are closely linked to their pathogenicity, and the lack of structural details have hindered our understanding of PD. Here, I present the first 3D structure of α-syn fibrils determined through solid- state NMR spectroscopy experiments validated with X-ray fiber diffraction and electron microscopy mass per-length measurements. The fibril form presented here is shown to be pathologically active in neuronal cell culture. Further, the structure is validated by comparison of the structure to three of the early-onset PD mutants, A30P, E46K, and A53T, and show that the changes in the SSNMR spectra induced by the mutation are consistent with the structural features present in the fibril, most notably the E46K mutant disrupting a stabilizing salt-bridge within the fibril core. Additionally, I show that non- native cysteine mutants of T33C, V48C, T59C, A85C, and Q99C further validate the structure. The results and studies presented here establish the first structure of α-syn fibrils and provide the experimental and computational basis for greatly increasing our understanding of PD, amyloid fibril structure, and advancing the techniques available for structural studies via SSNMR spectroscopy.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2017-12-01","The student, Marcus Tuttle, accepted the attached license on 2015-11-30 at 16:03.","The student, Marcus Tuttle, submitted this Dissertation for approval on 2015-11-30 at 16:23.","This Dissertation was approved for publication on 2015-12-01 at 12:09.","DSpace SAF Submission Ingestion Package generated from Vireo submission #8877 on 2016-03-02 at 14:13:31","Made available in DSpace on 2016-03-02T21:06:50Z (GMT). No. of bitstreams: 20 TUTTLE-DISSERTATION-2015.pdf: 25957302 bytes, checksum: e0ff2c528ec0f9585177d1b1ff5766e6 (MD5) Chapter_0a_Title_Page_SPACING.docx: 46703 bytes, checksum: 626a773119041caf0972cd994bb7b820 (MD5) Chapter_0b_Abstract_DONE.docx: 115185 bytes, checksum: 28042ff974493c824b35b8d1c027e05d (MD5) Chapter_0c_Acknowledgements_DONE.docx: 119424 bytes, checksum: 5f4b49133be9e5db06b20f28c354d30d (MD5) Chapter_0d_TableofContents_NEEDS_PAGENUMS.docx: 81655 bytes, checksum: 588b3ee9e1cc86614cf220f84c93f66c (MD5) Chapter_1_DONE.docx: 1240527 bytes, checksum: 500c637f4063b494a254b5cbc6b2e173 (MD5) Chapter_2_DONE.docx: 943435 bytes, checksum: a8b7dc731f90a4b090a07e2b3a655eb9 (MD5) Chapter_3_DONE.docx: 1840648 bytes, checksum: 029f682f63995ffbeeea2f070f4a950e (MD5) Chapter_4_DONE.docx: 867234 bytes, checksum: 492e11369a275b830feee8abf0fb1dcf (MD5) Chapter_5_DONE.docx: 10890464 bytes, checksum: 8baa4cee07aa6c6567a1f357e5d00361 (MD5) Chapter_6_DONE_18NOV15.docx: 1093414 bytes, checksum: 62d6ce9cfb681a7daf48c0e34238110c (MD5) Chapter_7_DONE.docx: 303105 bytes, checksum: 57a825c3a8302d5112b847056102a112 (MD5) Chapter_8_DONE.docx: 139574 bytes, checksum: 9ff6fff4015dc3c389f461fdf9c47cd2 (MD5) Chapter3_MiMB_License.pdf: 287085 bytes, checksum: 5d3cca17ca676720df9faa4d7f080746 (MD5) Chapter_1_Fig1.pdf: 574926 bytes, checksum: b39a6f39b6a4da71962ba9f3fb6174ec (MD5) Chapter_1_Fig2.pdf: 46094 bytes, checksum: 73bcec5a60f1aa95e345344264496cd4 (MD5) Chapter_3_Fig_9.pdf: 136509 bytes, checksum: 7f3c5bd517df90d85e5221baaa5cb85d (MD5) Chapter_6_Fig1.pdf: 668338 bytes, checksum: b2a33700b3e9951fdd41c29053bb1e55 (MD5) Glycerol_Labeling_Permission.pdf: 196227 bytes, checksum: 969a8f5f2e89a2d26e19ab5e5cd800ba (MD5) LICENSE.txt: 4210 bytes, checksum: 30579cafc7a0159016efb81c25b31cc5 (MD5) Previous issue date: 2015-12-01","Embargo set by: Seth Robbins for item 91419 Lift date: 2018-03-02T21:07:27Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Limited Restriction Lifted for Item 91419 on 2018-03-03T10:15:22Z."],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/2142/89216"],"dc:language":["en"],"dc:rights":["Copyright 2015 Marcus Dale Tuttle"],"dc:subject":["solid state nuclear magnetic resonance (SSNMR)","α-Synuclein","Parkinson's Disease","Nuclear magnetic resonance (NMR)","Structural Biology"],"dc:title":["Solid-state NMR studies of α-synuclein fibril structure"],"dc:type":["text"],"thesis:degree_discipline":["Chemistry"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:26:32Z"}