The Molecular Organization of the Fungal Prion HET-s in Its Amyloid Form

Wasmer, Christian; Schütz, Anne K.; Loquet, Antoine; Buhtz, Carolin; Greenwald, Jason; Riek, Roland; Böckmann, Anja; Meier, Beat H.

doi:10.1016/j.jmb.2009.09.015

Cited by 77 publications

(96 citation statements)

References 39 publications

(36 reference statements)

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“…Such methods have been successfully applied to probe motional disordered regions of membrane-associated proteins 11 and protein amyloids 12,13 and to reduce spectral complexity using molecular constructs of different size. 14 In the present work, we investigated the use of such methods to probe structural features of distinct topological regions of BamA.…”

mentioning

confidence: 99%

“…To characterize rigid and mobile protein segments, we acquired two-dimensional (2D) ( 13 C, 13 C) and ( 1 H, 13 C) correlation spectra employing dipolar and scalar magnetization transfer steps, respectively.…”

mentioning

confidence: 99%

“…11 Figure 1b shows a comparison between 2D 13 C- 13 C proton-driven spin diffusion (PDSD) spectra obtained using a short mixing time (40 ms) that were recorded on uniformly ( 13 C, 15 N)-labeled BamA precipitate (black) and reconstituted BamA(ΔP1-P4) (blue). Overall, the spectra display good dispersion and sensitivity and comparable spectral resolution.…”

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Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA

et al. 2011

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Multidomain proteins constitute a large part of prokaryotic and eukaryotic proteomes and play fundamental roles in various physiological processes. However, their structural characterization is challenging because of their large size and intrinsic flexibility. We show here that motional-filtered high-resolution solid-state NMR (ssNMR) experiments allow for the observation and structural analysis of very large multidomain membrane proteins that are characterized by different motional time scales. This approach was used to probe the folding of the 790-residue membrane protein BamA, which is the core component of the Escherichia coli outer membrane protein assembly machinery. A combination of dipolar-and scalar-based two-dimensional ssNMR experiments applied to two uniformly 13 C, 15 N-labeled BamA variants revealed characteristic secondary structure elements and distinct dynamics within the BamA transmembrane protein segment and the periplasmic POTRA domains. This approach hence provides a general strategy for collecting atomic-scale structural information on multidomain (membrane) proteins in a native-like environment.

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Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA

et al. 2011

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“…Some proteins can aggregate into amyloid-like structures when heat-denatured, such as in the poached-egg-white studies by Astbury et al (1935), and some proteins that have stable soluble folds can be transformed into an amyloid state in nonbiological conditions (Guijarro et al 1998;Chiti et al 1999). However, amyloid aggregation is highly amino acid sequence specific, as demonstrated by the structures discussed above (Nelson et al 2005;Nelson and Eisenberg 2006a;Sawaya et al 2007;Wasmer et al 2009). The essential involvement of side-chain interactions in the aggregation process is evident from the observed sequence-specific nature of amyloid aggregation (Tjernberg et al 2002;Zanuy and Nussinov 2003;Margittai and Langen 2006) and from the predictive power of the algorithms for aggregation propensities (Fernandez-Escamilla et al 2004;Trovato et al 2006;Tartaglia et al 2008).…”

Section: Can Any Protein Form An Amyloid?mentioning

confidence: 99%

The Three-Dimensional Structures of Amyloids

Riek

2016

Cold Spring Harb Perspect Biol

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Amyloids are highly ordered protein aggregates that are associated with both disease (including PrP prion, Alzheimer's, and Parkinson's) and biological function. The amyloid structure is composed of the cross-b-sheet entity, which is an almost indefinitely repeating twolayered intermolecular b-sheet motif. The three-dimensional (3D) structure is unique among protein folds because it folds only upon intermolecular contacts (for a folding to occur, only short sequences of amino acid residues are required), and the structure repeats itself at the atomic level (i.e., every 4.7 Å ). As a consequence of this structure, among others, it can grow by recruiting corresponding amyloid peptide/protein and thus has the capacity to be an infectious protein (i.e., a prion). Furthermore, its repetitiveness can translate what would be a nonspecific activity as monomer into a potent one through cooperativity. Because of these and other properties, the activities of amyloids are manifold and include peptide storage, template assistance, loss of function, gain of function, generation of toxicity, membrane binding, infectivity, and more. This review summarizes the structural nature of the cross-bsheet motif on the basis of a few high-resolution structural studies of amyloids in the context of potential biological activities.

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“…5 The HET-s protein displays a globular α-helical domain appended to a natively unfolded PFD that act together in a structural model wherein amyloid fibrils built from a cross-β core are decorated with globular domains. 30 The HET-s PFD structure based on ssNMR represents the first complete structure decrypted of an amyloid fibril. HET-s PFD fibrils assembled at neutral pH forms a left-handed β-solenoid, with each molecule forming two helical windings, and a highly ordered triangular cross-β compact hydrophobic core with three salt bridges and two asparagine ladders.…”

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Amyloids in solid-state nuclear magnetic resonance: potential causes of the usually low resolution

Espargaró¹,

Busquets²,

Estelrich³

et al. 2015

IJN

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Amyloids are non-crystalline and insoluble, which imply that the classical structural biology tools, ie, X-ray crystallography and solution nuclear magnetic resonance (NMR), are not suitable for their analysis. In the last years, solid-state NMR (ssNMR) has emerged as an alternative tool to decrypt the structural signatures of amyloid fibrils, providing major contributions to our understanding of molecular structures of amyloids such as β-amyloid peptide associated with Alzheimer's disease or fungal prions, among others. Despite this, the wide majority of amyloid fibrils display low resolution by ssNMR. Usually, this low resolution has been attributed to a high disorder or polymorphism of the fibrils, suggesting the existence of diverse elementary β-sheet structures. Here, we propose that a single β-sheet structure could be responsible for the broadening of the line widths in the ssNMR spectra. Although the fibrils and fibers consist of a single elementary structure, the angle of twist of each individual fibril in the mature fiber depends on the number of individual fibrils as well as the fibril arrangement in the final mature fiber. Thus, a wide range of angles of twist could be observed in the same amyloid sample. These twist variations involve changes in amino acid alignments that could be enough to limit the ssNMR resolution.

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The Molecular Organization of the Fungal Prion HET-s in Its Amyloid Form

Cited by 77 publications

References 39 publications

Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA

Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA

The Three-Dimensional Structures of Amyloids

Amyloids in solid-state nuclear magnetic resonance: potential causes of the usually low resolution

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