Prion Fibrils of Ure2p Assembled under Physiological Conditions Contain Highly Ordered, Natively Folded Modules

Loquet, Antoine; Bousset, Luc; Gardiennet, Carole; Sourigues, Yannick; Wasmer, Christian; Habenstein, Birgit; Schütz, Anne K.; Meier, Beat H.; Melki, Ronald; Böckmann, Anja

doi:10.1016/j.jmb.2009.09.016

Cited by 67 publications

(100 citation statements)

References 49 publications

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“…Such observations may not be contradictory, if the native conformations occur in non-amyloidogenic domains that remain attached to the amyloid itself (7,8). However, native interactions could well be preserved within the fibril core for proteins that natively contain ␤-sheets, as proposed for ␤2m and transthyretin, for instance (9,10).…”

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confidence: 99%

Amyloid-like Fibrils from a Domain-swapping Protein Feature a Parallel, in-Register Conformation without Native-like Interactions

Hoop

Kodali

et al. 2011

Journal of Biological Chemistry

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The formation of amyloid-like fibrils is characteristic of various diseases, but the underlying mechanism and the factors that determine whether, when, and how proteins form amyloid, remain uncertain. Certain mechanisms have been proposed based on the three-dimensional or runaway domain swapping, inspired by the fact that some proteins show an apparent correlation between the ability to form domain-swapped dimers and a tendency to form fibrillar aggregates. Intramolecular ␤-sheet contacts present in the monomeric state could constitute intermolecular ␤-sheets in the dimeric and fibrillar states. One example is an amyloid-forming mutant of the immunoglobulin binding domain B1 of streptococcal protein G, which in its native conformation consists of a four-stranded ␤-sheet and one ␣-helix. Under native conditions this mutant adopts a domainswapped dimer, and it also forms amyloid-like fibrils, seemingly in correlation to its domain-swapping ability. We employ magic angle spinning solid-state NMR and other methods to examine key structural features of these fibrils. Our results reveal a highly rigid fibril structure that lacks mobile domains and indicate a parallel in-register ␤-sheet structure and a general loss of native conformation within the mature fibrils. This observation contrasts with predictions that native structure, and in particular intermolecular ␤-strand interactions seen in the dimeric state, may be preserved in "domain-swapping" fibrils. We discuss these observations in light of recent work on related amyloidforming proteins that have been argued to follow similar mechanisms and how this may have implications for the role of domain-swapping propensities for amyloid formation.Amyloid fibril formation is characteristic of a variety of human disorders, including Huntington and Alzheimer diseases (1, 2). In amyloid-related diseases, one or more proteins are found in fibrillar aggregates, in a non-native, highly ␤-sheetrich conformation. Depending on the disease context, the propensity for amyloid formation may be traced to mutations and cleavage events, combined with poorly understood external triggers. Many proteins can also be made to form amyloid fibrils in vitro. Intriguingly, there are many accounts of proteins that form amyloid-like fibrils that are not associated with pathologies (2). Whether disease-related or not, amyloid fibrils share key biochemical and biophysical characteristics, suggesting common structural features. Understanding the fibril formation pathway is of interest not only as there may be a correlation between disease onset and protein aggregation, but also because transient oligomeric precursors may act as toxic species. However, a lack of high resolution structures of most fibrils and their precursors limits our knowledge of the mechanism of formation.One seemingly common structural motif for amyloid fibrils is an in-register parallel (IP) 3 assembly into pleated ␤-sheets, stabilized by backbone-to-backbone hydrogen bonding, combined with favorable side-chain interactions (e....

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confidence: 99%

Amyloid-like Fibrils from a Domain-swapping Protein Feature a Parallel, in-Register Conformation without Native-like Interactions

Hoop

Kodali

et al. 2011

Journal of Biological Chemistry

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“…39,[55][56][57] This globular domain retains its native structure in the fibrillar form of the protein. [58][59][60] While the C-terminal domain alone can support the nitrogen regulation function of Ure2p (and complement a Δure2 allele), the the light of recent findings on Sup35p and Ure2p assembly into infectious fibrils from our laboratory and others.…”

Section: Yeast Prions Assembly and Propagationmentioning

confidence: 90%

“…Firstly, the C-terminal domains could interact with the N-terminal domains in the soluble form of the proteins. 7,48,60,76 Such an interaction should impact the ensemble presence of the N-terminal domain appears to be required for tighter nitrogen catabolite repression under normal Ure2p expression levels. 39,61,62 Ure2p displays glutathione peroxidase and glutaredoxin activities, in both the soluble and fibrillar state, and therefore contributes to cell protection against heavy metal ions and oxidant toxicity.…”

Section: The [Psi + ] and [Ure3] Prionsmentioning

confidence: 99%

“…Two forms of Ure2p fibrils have been reported: one with an α-helical content, an X-ray fiber diffraction, solid state NMR, H/D exchange and proteolytic patterns that are not those of a classical amyloids; 58,60,87,89 another that has the typical characteristics of amyloids. 85,90 The latter form may be the consequence of fibril dehydration.…”

Section: The [Psi + ] and [Ure3] Prionsmentioning

confidence: 99%

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Yeast prions assembly and propagation: Contributions of the prion and non-prion moieties and the nature of assemblies

Kabani

Melki

2011

Prion

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“…For the solid‐state NMR investigation, ANSII (either free or its MenC conjugate) was freeze‐dried, packed into the magic‐angle spinning (MAS) rotors, then rehydrated to restore the resolution 30, 34, 35, 36. The spectra collected on the pelleted ANSII‐MenC conjugate (Figure 2) are largely superimposable to those of the native and PEGylated forms of the enzyme (see Figure S2 in the Supporting Information) making possible the assessment of the preservation of the protein fold after conjugation with the MenC polysaccharide 37, 38, 39, 40. The resonances of the protein residues were easily reassigned starting from the assignment of the native form of ANSII, by using a carbon‐detection approach (Figure S3) 41.…”

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Characterization of the Conjugation Pattern in Large Polysaccharide–Protein Conjugates by NMR Spectroscopy

et al. 2017

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Carbohydrate‐based vaccines are among the safest and most effective vaccines and represent potent tools for prevention of life‐threatening bacterial infectious diseases, like meningitis and pneumonia. The chemical conjugation of a weak antigen to protein as a source of T‐cell epitopes generates a glycoconjugate vaccine that results more immunogenic. Several methods have been used so far to characterize the resulting polysaccharide–protein conjugates. However, a reduced number of methodologies has been proposed for measuring the degree of saccharide conjugation at the possible protein sites. Here we show that detailed information on large proteins conjugated with large polysaccharides can be achieved by a combination of solution and solid‐state NMR spectroscopy. As a test case, a large protein assembly, l‐asparaginase II, has been conjugated with Neisseria meningitidis serogroup C capsular polysaccharide and the pattern and degree of conjugation were determined.

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Prion Fibrils of Ure2p Assembled under Physiological Conditions Contain Highly Ordered, Natively Folded Modules

Cited by 67 publications

References 49 publications

Amyloid-like Fibrils from a Domain-swapping Protein Feature a Parallel, in-Register Conformation without Native-like Interactions

Amyloid-like Fibrils from a Domain-swapping Protein Feature a Parallel, in-Register Conformation without Native-like Interactions

Yeast prions assembly and propagation: Contributions of the prion and non-prion moieties and the nature of assemblies

Characterization of the Conjugation Pattern in Large Polysaccharide–Protein Conjugates by NMR Spectroscopy

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