NMR paper Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register.

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[NMR paper] Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register.

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NMR processing:

MDD

NMR assignment:

Backbone:

Side-chains:

NOEs:

Structure from NMR restraints:

Ab initio:

Fragment-based:

Rosetta-NMR (Robetta)

Template-based:

GeNMR

I-TASSER

Refinement:

Amber

Structure from chemical shifts:

Fragment-based:

Homology-based:

Torsion angles from chemical shifts:

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TALOS

Promega- Proline

Secondary structure from chemical shifts:

CSI (via RCI server)

TALOS

MICS caps, β-turns

d2D

PECAN

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RCI

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HADDOCK

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Methyl S2

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ArShift- Aromatic

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Shifty

Camcoil

Poulsen_rc_CS

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Format conversion & validation:

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03-24-2017, 10:14 PM

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Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register.

Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register.

Related Articles Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register.

Proc Natl Acad Sci U S A. 2017 Mar 22;:

Authors: Frederick KK, Michaelis VK, Caporini MA, Andreas LB, Debelouchina GT, Griffin RG, Lindquist S

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 ?-helical arrangement, whereas others suggest a parallel in-register 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.

PMID: 28330994 [PubMed - as supplied by publisher]

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