Conserved features of intermediates in amyloid assembly determine their benign or toxic states
Some amyloid-forming polypeptides are associated with devastating human diseases and others provide important biological functions. For both, oligomeric intermediates appear during amyloid assembly. Currently we have few tools for characterizing these conformationally labile intermediates and discer...
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| Format: | Article |
| Language: | en_US |
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National Academy of Sciences
2013
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| Online Access: | http://hdl.handle.net/1721.1/77145 https://orcid.org/0000-0003-1307-882X |
| _version_ | 1826209947161985024 |
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| author | Krishnan, Rajaraman Goodman, Jessica L. Mukhopadhyay, Samrat Pacheco, Chris D. Lemke, Edward A. Deniz, Ashok A. Lindquist, Susan |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Krishnan, Rajaraman Goodman, Jessica L. Mukhopadhyay, Samrat Pacheco, Chris D. Lemke, Edward A. Deniz, Ashok A. Lindquist, Susan |
| author_sort | Krishnan, Rajaraman |
| collection | MIT |
| description | Some amyloid-forming polypeptides are associated with devastating human diseases and others provide important biological functions. For both, oligomeric intermediates appear during amyloid assembly. Currently we have few tools for characterizing these conformationally labile intermediates and discerning what governs their benign versus toxic states. Here, we examine intermediates in the assembly of a normal, functional amyloid, the prion-determining region of yeast Sup35 (NM). During assembly, NM formed a variety of oligomers with different sizes and conformation-specific antibody reactivities. Earlier oligomers were less compact and reacted with the conformational antibody A11. More mature oligomers were more compact and reacted with conformational antibody OC. We found we could arrest NM in either of these two distinct oligomeric states with small molecules or crosslinking. The A11-reactive oligomers were more hydrophobic (as measured by Nile Red binding) and were highly toxic to neuronal cells, while OC-reactive oligomers were less hydrophobic and were not toxic. The A11 and OC antibodies were originally raised against oligomers of Aβ, an amyloidogenic peptide implicated in Alzheimer’s disease (AD) that is completely unrelated to NM in sequence. Thus, this natural yeast prion samples two conformational states similar to those sampled by Aβ, and when assembly stalls at one of these two states, but not the other, it becomes extremely toxic. Our results have implications for selective pressures operating on the evolution of amyloid folds across a billion years of evolution. Understanding the features that govern such conformational transitions will shed light on human disease and evolution alike. |
| first_indexed | 2024-09-23T14:36:46Z |
| format | Article |
| id | mit-1721.1/77145 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:36:46Z |
| publishDate | 2013 |
| publisher | National Academy of Sciences |
| record_format | dspace |
| spelling | mit-1721.1/771452022-10-01T21:47:39Z Conserved features of intermediates in amyloid assembly determine their benign or toxic states Krishnan, Rajaraman Goodman, Jessica L. Mukhopadhyay, Samrat Pacheco, Chris D. Lemke, Edward A. Deniz, Ashok A. Lindquist, Susan Massachusetts Institute of Technology. Department of Biology Lindquist, Susan Some amyloid-forming polypeptides are associated with devastating human diseases and others provide important biological functions. For both, oligomeric intermediates appear during amyloid assembly. Currently we have few tools for characterizing these conformationally labile intermediates and discerning what governs their benign versus toxic states. Here, we examine intermediates in the assembly of a normal, functional amyloid, the prion-determining region of yeast Sup35 (NM). During assembly, NM formed a variety of oligomers with different sizes and conformation-specific antibody reactivities. Earlier oligomers were less compact and reacted with the conformational antibody A11. More mature oligomers were more compact and reacted with conformational antibody OC. We found we could arrest NM in either of these two distinct oligomeric states with small molecules or crosslinking. The A11-reactive oligomers were more hydrophobic (as measured by Nile Red binding) and were highly toxic to neuronal cells, while OC-reactive oligomers were less hydrophobic and were not toxic. The A11 and OC antibodies were originally raised against oligomers of Aβ, an amyloidogenic peptide implicated in Alzheimer’s disease (AD) that is completely unrelated to NM in sequence. Thus, this natural yeast prion samples two conformational states similar to those sampled by Aβ, and when assembly stalls at one of these two states, but not the other, it becomes extremely toxic. Our results have implications for selective pressures operating on the evolution of amyloid folds across a billion years of evolution. Understanding the features that govern such conformational transitions will shed light on human disease and evolution alike. 2013-02-15T15:52:23Z 2013-02-15T15:52:23Z 2012-07 2012-06 Article http://purl.org/eprint/type/JournalArticle 0027-8424 1538-4357 http://hdl.handle.net/1721.1/77145 Krishnan, R. et al. “Conserved Features of Intermediates in Amyloid Assembly Determine Their Benign or Toxic States.” Proceedings of the National Academy of Sciences 109.28 (2012): 11172–11177. https://orcid.org/0000-0003-1307-882X en_US http://dx.doi.org/10.1073/pnas.1209527109 Proceedings of the National Academy of Sciences Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences PNAS |
| spellingShingle | Krishnan, Rajaraman Goodman, Jessica L. Mukhopadhyay, Samrat Pacheco, Chris D. Lemke, Edward A. Deniz, Ashok A. Lindquist, Susan Conserved features of intermediates in amyloid assembly determine their benign or toxic states |
| title | Conserved features of intermediates in amyloid assembly determine their benign or toxic states |
| title_full | Conserved features of intermediates in amyloid assembly determine their benign or toxic states |
| title_fullStr | Conserved features of intermediates in amyloid assembly determine their benign or toxic states |
| title_full_unstemmed | Conserved features of intermediates in amyloid assembly determine their benign or toxic states |
| title_short | Conserved features of intermediates in amyloid assembly determine their benign or toxic states |
| title_sort | conserved features of intermediates in amyloid assembly determine their benign or toxic states |
| url | http://hdl.handle.net/1721.1/77145 https://orcid.org/0000-0003-1307-882X |
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