Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register
Name
Frederick-2017-Combining DNP NMR with segmenta.pdf
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Author(s)
Frederick, Kendra K.
Caporini, Marc A.
Michaelis, Vladimir K.
Andreas, Loren
Lindquist, Susan
Debelouchina, Galia Tzvetanova
Griffin, Robert Guy
Date Issued
March 2017
Journal
Proceedings of the National Academy of Sciences
Publisher
Proceedings of the National Academy of Sciences
Citation
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
Version
Final published version
Abstract
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
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemistry
Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology)
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DOI of Published Version
https://doi.org/10.1073/PNAS.1619051114
