Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register
The yeast prion protein Sup35NM is a self-propagating amyloid. Despite intense study, there is no consensus on the organization of monomers within Sup35NM fibrils. Some studies point to a â-helical arrangement, whereas others suggest a parallel inregister organization. Intermolecular contacts are of...
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Proceedings of the National Academy of Sciences
2017
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| Online Access: | http://hdl.handle.net/1721.1/112944 https://orcid.org/0000-0002-6708-7660 https://orcid.org/0000-0003-1307-882X https://orcid.org/0000-0003-1589-832X |
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| author | Frederick, Kendra K. Caporini, Marc A. Michaelis, Vladimir K. Andreas, Loren Lindquist, Susan Debelouchina, Galia Tzvetanova Griffin, Robert Guy |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Frederick, Kendra K. Caporini, Marc A. Michaelis, Vladimir K. Andreas, Loren Lindquist, Susan Debelouchina, Galia Tzvetanova Griffin, Robert Guy |
| author_sort | Frederick, Kendra K. |
| collection | MIT |
| description | The yeast prion protein Sup35NM is a self-propagating amyloid. Despite intense study, there is no consensus on the organization of monomers within Sup35NM fibrils. Some studies point to a â-helical arrangement, whereas others suggest a parallel inregister organization. Intermolecular contacts are often determined by experiments that probe long-range heteronuclear contacts for fibrils templated from a 1:1 mixture of 13 C- and 15 N-labeled monomers. However, for Sup35NM, like many large proteins, chemical shift degeneracy limits the usefulness of this approach. Segmental and specific isotopic labeling reduce degeneracy, but experiments to measure long-range interactions are often too insensitive. To limit degeneracy and increase experimental sensitivity, we combined specific and segmental isotopic labeling schemes with dynamic nuclear polarization (DNP) NMR. Using this combination, we examined an amyloid form of Sup35NM that does not have a parallel in-register structure. The combination of a small number of specific labels with DNP NMR enables determination of architectural information about polymeric protein systems. Keyword: [PSI+] prion; solid-state NMR; amyloid; Sup35; dynamic nuclear polarization |
| first_indexed | 2024-09-23T13:11:59Z |
| format | Article |
| id | mit-1721.1/112944 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T13:11:59Z |
| publishDate | 2017 |
| publisher | Proceedings of the National Academy of Sciences |
| record_format | dspace |
| spelling | mit-1721.1/1129442022-09-28T12:36:34Z Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register Frederick, Kendra K. Caporini, Marc A. Michaelis, Vladimir K. Andreas, Loren Lindquist, Susan Debelouchina, Galia Tzvetanova Griffin, Robert Guy Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology) Michaelis, Vladimir K. Andreas, Loren Lindquist, Susan Debelouchina, Galia Tzvetanova Griffin, Robert Guy The yeast prion protein Sup35NM is a self-propagating amyloid. Despite intense study, there is no consensus on the organization of monomers within Sup35NM fibrils. Some studies point to a â-helical arrangement, whereas others suggest a parallel inregister organization. Intermolecular contacts are often determined by experiments that probe long-range heteronuclear contacts for fibrils templated from a 1:1 mixture of 13 C- and 15 N-labeled monomers. However, for Sup35NM, like many large proteins, chemical shift degeneracy limits the usefulness of this approach. Segmental and specific isotopic labeling reduce degeneracy, but experiments to measure long-range interactions are often too insensitive. To limit degeneracy and increase experimental sensitivity, we combined specific and segmental isotopic labeling schemes with dynamic nuclear polarization (DNP) NMR. Using this combination, we examined an amyloid form of Sup35NM that does not have a parallel in-register structure. The combination of a small number of specific labels with DNP NMR enables determination of architectural information about polymeric protein systems. Keyword: [PSI+] prion; solid-state NMR; amyloid; Sup35; dynamic nuclear polarization National Institutes of Health (U.S.) (Grants GM-025874) National Institutes of Health (U.S.) (Grants EB-003151) National Institutes of Health (U.S.) (Grants EB-002804) National Institutes of Health (U.S.) (Grants EB-002026) 2017-12-22T20:47:11Z 2017-12-22T20:47:11Z 2017-03 2016-11 2017-12-22T17:23:06Z Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/112944 Frederick, Kendra K., et al. “Combining DNP NMR with Segmental and Specific Labeling to Study a Yeast Prion Protein Strain That Is Not Parallel in-Register.” Proceedings of the National Academy of Sciences, vol. 114, no. 14, Apr. 2017, pp. 3642–47. © 2017 National Academy of Sciences https://orcid.org/0000-0002-6708-7660 https://orcid.org/0000-0003-1307-882X https://orcid.org/0000-0003-1589-832X http://dx.doi.org/10.1073/PNAS.1619051114 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 Proceedings of the National Academy of Sciences PNAS |
| spellingShingle | Frederick, Kendra K. Caporini, Marc A. Michaelis, Vladimir K. Andreas, Loren Lindquist, Susan Debelouchina, Galia Tzvetanova Griffin, Robert Guy Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register |
| title | Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register |
| title_full | Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register |
| title_fullStr | Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register |
| title_full_unstemmed | Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register |
| title_short | Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register |
| title_sort | combining dnp nmr with segmental and specific labeling to study a yeast prion protein strain that is not parallel in register |
| url | http://hdl.handle.net/1721.1/112944 https://orcid.org/0000-0002-6708-7660 https://orcid.org/0000-0003-1307-882X https://orcid.org/0000-0003-1589-832X |
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