Observation of intermediate states of the human prion protein by high pressure NMR spectroscopy

@article{Kachel2006ObservationOI,
  title={Observation of intermediate states of the human prion protein by high pressure NMR spectroscopy},
  author={Norman Kachel and Werner Kremer and Ralph Zahn and Hans Robert Kalbitzer},
  journal={BMC Structural Biology},
  year={2006},
  volume={6},
  pages={16 - 16}
}
BackgroundPrions as causative agents of transmissible spongiform encephalopathies (TSEs) in humans and animals are composed of the infectious isomer, PrPSc, of the cellular prion protein, PrPC. The conversion and thus the propensity of PrPC to adopt alternative folds leads to the species-specific propagation of the disease. High pressure is a powerful tool to study the physico-chemical properties of proteins as well as the dynamics and structure of folding intermediates.ResultsConformational… Expand
Differential stability of the bovine prion protein upon urea unfolding
TLDR
High‐resolution NMR spectroscopy is used to characterize the stability and structure of bovine recombinant PrPC during unfolding with the denaturant urea, suggesting that the dissociation of the native β‐sheet of PrPC is a primary step in the urea‐induced unfolding process, while strong hydrophobic interactions between helices α1 and α2, and between α2 and α3, stabilize these regions even at very high concentrations of urea. Expand
Influence of pH on the human prion protein: insights into the early steps of misfolding.
TLDR
Detailed molecular-dynamics simulations of the recombinant PrP protein in water at three different pH regimes provide a molecular basis for the initial steps in the misfolding process and are consistent with experimental data. Expand
Structural transitions in full-length human prion protein detected by xenon as probe and spin labeling of the N-terminal domain
TLDR
Xenon bound PrP was modelled by restraint molecular dynamics and the individual microscopic and macroscopic dissociation constants could be derived by fitting the data to a model including a dynamic opening and closing of the cavities. Expand
Partially Unfolded Forms of the Prion Protein Populated under Misfolding-promoting Conditions
TLDR
Hydrogen-deuterium exchange in conjunction with mass spectrometry and nuclear magnetic resonance spectroscopy were used for the structural and energetic characterization of the N state of the full-length mouse prion protein, moPrP, under conditions that favor misfolding. Expand
Structural and hydration properties of the partially unfolded states of the prion protein.
TLDR
The free energy surface of the C-terminal globular domain of the PrP is constructed from enhanced sampling of replica exchange molecular dynamics to gain insights into possible aggregation-prone intermediate states. Expand
Prion protein and its conformational conversion: a structural perspective.
TLDR
Recent advances in understanding biophysical and biochemical aspects of prion diseases are described, with a special focus on structural underpinnings ofPrP protein conversion, the structural basis of prions strains, and generation ofPrion infectivity in vitro from bacterially-expressed recombinant PrP. Expand
Energy landscape of the prion protein helix 1 probed by metadynamics and NMR.
TLDR
The combined approach allowed us to dissect the factors that govern the conformational states of PrP-H1 in solution, and the implications of these factors for prion protein misfolding and aggregation. Expand
Reversible monomer-oligomer transition in human prion protein
TLDR
The results strongly suggest that the oligomeric form PrPoligo is in dynamic equilibrium with the monomeric forms via PrPC*, namely huPrPC ⇄ hu PrPC* ⇄huPrPoligo. Expand
Interplay of buried histidine protonation and protein stability in prion misfolding
TLDR
Analysis of the data indicates that protonation of the buried histidine destabilizes both PrP variants, but produces a more drastic effect in the less stable GHaPrP. Expand
Conformational Properties of β-PrP*
TLDR
It is found that a disulfide-reduced fragment of human PrP spanning residues 91–231 under acidic conditions, which similarly to PrPSc, is a potent inhibitor of the 26 S proteasome, assembles into soluble oligomers that have significant β-sheet content. Expand
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