The Effect of Disease-associated Mutations on the Folding Pathway of Human Prion Protein

Apetri, Adrian; Surewicz, Krystyna; Surewicz, Witold K.

doi:10.1074/jbc.m313581200

Cited by 146 publications

(154 citation statements)

References 58 publications

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“…Previous kinetic studies of PrP folding have shown that at low temperature (5°C), the human protein folds by way of a populated intermediate (14)(15)(16). The rate-limiting conversion of this species into the native state occurs at Ϸ1,400 s Ϫ1 , some 14-fold slower than the rate-limiting step at 25°C measured in our current study.…”

Section: Discussioncontrasting

confidence: 39%

“…Because we cannot perform perturbation experiments at low denaturant concentrations, it is not possible to detect the presence of such an unstable intermediate by using our higher-temperature data. A major objective of the work of Surewicz and colleagues was to investigate the effects of naturally occurring mutations in PrP that lead to inherited prion disease (15). The broad conclusion was that 2/3 of these stabilized the intermediate, but folding rates were essentially the same.…”

Section: Discussionmentioning

confidence: 99%

“…However, an inherent limitation of the hydrogen-exchange technique is that it provides information only for those regions that display measurable protection in the folded state. To extend our analysis, we compared the conformational dynamics of both mutants through the use of 15 The NMR data thus indicate that the I184V mutation has no effect on the native state. We are therefore left with the conclusion that the polypeptide has 2 potential trajectories along which it can fold.…”

Section: Discussionmentioning

confidence: 99%

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Folding kinetics of the human prion protein probed by temperature jump

Hart

Hosszu

Trevitt

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Temperature-jump perturbation was used to examine the relaxation kinetics of folding of the human prion protein. Measured rates were very fast (Ϸ3,000 s ؊1 ), with the extrapolated folding rate constant at Ϸ20°C in physiological conditions reaching 20,000 s ؊1 . By a mutational analysis of core residues, we found that only 2, on the interface of helices 2 and 3, have significant -values in the transition state. Interestingly, a mutation sandwiched between the above 2 residues on the helix-helix contact interface had very little effect on the overall free energy of folding but led to the formation of a monomeric misfolded state, which had to unfold to acquire the native PrP C conformation. Another mutation that led to a marked destabilization of the native fold also formed a misfolded intermediate, but this was aggregation-prone despite the native state of this mutant being soluble. Taken together, the data imply that this fast-folding protein has a transition state that is not compact (m value analysis gives a ␤t value of only 0.3) but contains a developing nucleus between helices 2 and 3. The fact that a mutation in this nucleus had a negligible effect on stability but still led to formation of aberrant conformations during folding implies an easily perturbed folding mechanism. It is notable that in inherited forms of human prion disease, where point mutations produce a lethal dominant condition, 20 of the 33 amino acid replacements occur in the helix-2/3 sequence. equilibrium perturbation ͉ -value analysis ͉ protein folding

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Section: Discussioncontrasting

confidence: 39%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Folding kinetics of the human prion protein probed by temperature jump

Hart

Hosszu

Trevitt

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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“…Remarkably, the vast majority of point mutations associated with these disorders map to the region found herein to undergo a transition to ␤-structure (4). Thus, these mutations are well positioned to cause structural and/or thermodynamic perturbation of this critical region in PrP C (40), thereby facilitating the spontaneous conversion to the PrP Sc isoform. The present structural model may also offer a rationale for baffling findings that the pattern of glycosylation (i.e., the ratio of diglycosylated, monoglycosylated, and unglycosylated PrP) is one of the determinants of prion strain diversity, being fatefully propagated upon transmission (2,4).…”

Section: Discussionmentioning

confidence: 99%

Molecular architecture of human prion protein amyloid: A parallel, in-register β-structure

Cobb

Sönnichsen

Mchaourab³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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342

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Transmissible spongiform encephalopathies (TSEs) represent a group of fatal neurodegenerative diseases that are associated with conformational conversion of the normally monomeric and ␣-helical prion protein, PrP C , to the ␤-sheet-rich PrP Sc . This latter conformer is believed to constitute the main component of the infectious TSE agent. In contrast to high-resolution data for the PrP C monomer, structures of the pathogenic PrP Sc or synthetic PrP Sc -like aggregates remain elusive. Here we have used sitedirected spin labeling and EPR spectroscopy to probe the molecular architecture of the recombinant PrP amyloid, a misfolded form recently reported to induce transmissible disease in mice overexpressing an N-terminally truncated form of PrP C . Our data show that, in contrast to earlier, largely theoretical models, the conformational conversion of PrP C involves major refolding of the C-terminal ␣-helical region. The core of the amyloid maps to C-terminal residues from Ϸ160 -220, and these residues form single-molecule layers that stack on top of one another with parallel, in-register alignment of ␤-strands. This structural insight has important implications for understanding the molecular basis of prion propagation, as well as hereditary prion diseases, most of which are associated with point mutations in the region found to undergo a refolding to ␤-structure.EPR ͉ spin labeling ͉ transmissible spongiform encephalopathy

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“…Experiments on mouse PrP mutants indicate that thermodynamic stability of PrP is significantly reduced for T183A and F198S and somewhat reduced for V180I [136]. For V180I, V210 and F198S PrP, it was found that a folding intermediate increased in the population relative to the native fold, further indicating that these mutations cause instability [89,151]. Studies of V203I PrP are limited, but may suggest an effect on stability as well [152].…”

Section: Mutations In the Hydrophobic Corementioning

confidence: 99%