Amyloid Formation by Recombinant Full-length Prion Proteins in Phospholipid Bicelle Solutions

Lührs, Thorsten; Zahn, Ralph; Wüthrich, Kurt

doi:10.1016/j.jmb.2006.01.016

Cited by 87 publications

(87 citation statements)

References 54 publications

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“…1 red) showed a typical β-sheet pattern, indicating that the α-rich form of human PrP C turned into the "β-rich form". The CD-spectra a very similar to the spectra presented by Lührs et al, 14 for native monomeric protein and an oligomeric form called PrP β that was induced by heat treatment in the presence of phospholipids.…”

Section: Resultssupporting

confidence: 82%

Reversible monomer-oligomer transition in human prion protein

Sasaki

Gaikwad

Hashiguchi

et al. 2008

Prion

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The structure and the dissociation reaction of oligomers PrP oligo from reduced human prion huPrP C (23-231) have been studied by 1 H-NMR and tryptophan fluorescence spectroscopy at varying pressure, along with circular dichroism and atomic force microscopy. The 1 H-NMR and fluorescence spectral feature of the oligomer is consistent with the notion that the N-terminal residues including all seven Trp residues, are free and mobile, while residues 105~210, comprising the AGAAAAGA motif and S1-Loop-HelixA-Loop-S2-Loop-HelixC, are engaged in intra-and/ or inter-molecular interactions. By increasing pressure to 200 MPa, the oligomers tend to dissociate into monomers which may be identified with PrP C* , a rare metastable form of PrP C stabilized at high pressure (Kachel et al., BMC Struct Biol 6:16). The results strongly suggest that the oligomeric form PrP oligo is in dynamic equilibrium with the monomeric forms via PrP C* , namely huPrP C  huPrP C*  huPrP oligo .

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Section: Resultssupporting

confidence: 82%

Reversible monomer-oligomer transition in human prion protein

Sasaki

Gaikwad

Hashiguchi

et al. 2008

Prion

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“…5). In the absence of denaturant the CD response is dominated by a ␤-sheet-rich species, and is similar to a soluble oligomeric form of PrP that was induced by heat treatment of ␣-PrP in the presence of phospholipids (31). At 5 M urea, however, the CD spectrum corresponds to a largely unfolded species, in line with the NMR data at high urea concentrations.…”

Section: Resultssupporting

confidence: 80%

“…This result is intriguing, suggesting a role for the pre- dominantly unstructured peptide segment 23-90 in the conformational transition to a distinct oligomeric species. This is not without precedent, as Lührs et al (31) have observed that peptide segment 105-120 is required for successful conversion of ␣-PrP into a soluble oligomeric species, which displays similar CD spectra to that observed for ␤-PrP, and there is evidence for some residues of the unfolded region of PrP transiently contacting the folded region of PrP (27,76).…”

Section: Discussionmentioning

confidence: 99%

“…The majority of these latter species have shown a high propensity for aggregation, although not all are on-pathway to the formation of amyloid. Many of these non-native states also display some of the characteristics of PrP Sc , such as increased ␤-sheet content, protease resistance, and a propensity for oligomerization (28,29,31) and some have been claimed to be associated with the disease process (34).…”

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confidence: 99%

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Conformational Properties of β-PrP

Hosszu

Trevitt

Jones

et al. 2009

Journal of Biological Chemistry

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Prion propagation involves a conformational transition of the cellular form of prion protein (PrP C ) to a disease-specific isomer (PrP Sc ), shifting from a predominantly ␣-helical conformation to one dominated by ␤-sheet structure. This conformational transition is of critical importance in understanding the molecular basis for prion disease. Here, we elucidate the conformational properties of a disulfide-reduced fragment of human PrP spanning residues 91-231 under acidic conditions, using a combination of heteronuclear NMR, analytical ultracentrifugation, and circular dichroism. We find that this form of the protein, which similarly to PrP Sc , is a potent inhibitor of the 26 S proteasome, assembles into soluble oligomers that have significant ␤-sheet content. The monomeric precursor to these oligomers exhibits many of the characteristics of a molten globule intermediate with some helical character in regions that form helices I and III in the PrP C conformation, whereas helix II exhibits little evidence for adopting a helical conformation, suggesting that this region is a likely source of interaction within the initial phases of the transformation to a ␤-rich conformation. This precursor state is almost as compact as the folded PrP C structure and, as it assembles, only residues 126 -227 are immobilized within the oligomeric structure, leaving the remainder in a mobile, random-coil state.

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“…The recombinant amyloidogenic prion protein, bPrP-β, is generated through chemical treatment of recombinant bPrP which enhances the formation of the β-sheet secondary structure in the C-terminal region. 37 The N-terminus of PrP (amino acids 23-120 in mPrP) is a flexible non-structural region and the C-terminus of PrP is preserved in the globular three-dimensional structure. 38 It is known that the N-terminal region of PrP interacts with nucleic acids in non-specific and/or specific manner.…”

Section: Introductionmentioning

confidence: 99%

Anti-bovine Prion protein RNA aptamer containing tandem GGA repeat interacts both with recombinant bovine prion protein and its β isoform with high affinity

Murakami

Nishikawa

Noda

et al. 2008

Prion

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In order to obtain RNA aptamers against bovine prion protein (bPrP), we carried out in vitro selection from RNA pools containing a 55-nucleotide randomized region to target recombinant bPrP. Most of obtained aptamers contained conserved GGA tandem repeats (GGA) 4 and aptamer #1 (apt #1) showed a high affinity for both bPrP and its β isoform (bPrP-β). The sequence of apt #1 suggested that it would have a G-quadruplex structure, which was confirmed using CD spectra in titration with KCl. A mutagenic study of this conserved region, and competitive assays, showed that the conserved (GGA) 4 sequence is important for specific binding to bPrP and bPrP-β. Following 5'-biotinylation, aptamer #1 specifically detected PrP c in bovine brain homogenate in a Northwestern blotting assay. Protein deletion mutant analysis showed that the bPrP aptamer binds within 25-131 of the bPrP sequence. Interestingly, the minimized aptamer #1 (17 nt) showed greater binding to bPrP and bPrP-β as compared to apt #1. This minimized form of aptamer #1 may therefore be useful in the detection of bPrP, diagnosis of prion disease, enrichment of bPr and ultimately in gaining a better understanding of prion diseases.

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Amyloid Formation by Recombinant Full-length Prion Proteins in Phospholipid Bicelle Solutions

Cited by 87 publications

References 54 publications

Reversible monomer-oligomer transition in human prion protein

Reversible monomer-oligomer transition in human prion protein

Conformational Properties of β-PrP

Anti-bovine Prion protein RNA aptamer containing tandem GGA repeat interacts both with recombinant bovine prion protein and its β isoform with high affinity

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