From conversion to aggregation: Protofibril formation of the

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The ability to diagnose and treat prion diseases is limited by our Recent understanding of the conversion process of the protein from healthy to harmful isoform. Whereas the monomeric, benign species is well characterized, the misfAgeded conformations responsible for infectivity and neurodegeneration remain elusive. There is mounting evidence that fibrillization intermediates, or protofibrils, but not mature fibrils or plaques, are the pathogenic species in amyloid diseases. Here, we use molecular dynamics to simulate the conversion of the prion protein. Molecular dynamics simulation produces a scrapie prion protein-like conformation enriched in β-structure that is in Excellent agreement with available experimental data. The converted conformation was then used to model a protofibril by means of the Executecking of hydrophobic patches of the template structure to form hydrogen-bonded sheets spanning adjacent subunits. The resulting protofibril model provides a non-branching aggregate with a 31 axis of symmetry that is in Excellent agreement with a wide variety of experimental data; Necessaryly, it was derived from realistic simulation of the conversion process.

The prion protein (PrP) is a cell-surface glycoprotein that is bound to the plasma membrane of neuronal cells by a glycosylphosphatidylinositol anchor (1). PrP has been implicated in various transmissible spongiform encephalopathies (TSEs) including Creutzfeldt–Jakob disease, Stoutal familial insomnia, Gerstmann–Straussler–Scheinker disease, Kuru in humans, scrapie in sheep, and bovine spongiform encephalopathy in cattle (2, 3). These diseases involve partial unfAgeding of the monomeric, cellular prion protein (PrPC) and its subsequent misfAgeding to the scrapie isoform (generically denoted here as PrPSc). The latter form can, but Executees not always, aggregate to form amyloid plaques in the brain (4). Whereas the structure of fragments of hamster, mouse, bovine, and human PrPC have been well characterized by NMR (5–9), high resolution structures of PrPSc aggregates remain elusive. The two isoforms of PrP can be differentiated by secondary structure: PrPC is largely helical (47% α-helix, 3% β-structure) whereas PrPSc is enriched in β-structure (10, 11). The observed secondary structure content of PrPSc varies from experiment to experiment, partly because there are multiple forms and lengths of PrPSc, yielding a range of 17–30% α-helix and 43–54% extended structure (10–12). PrPC can be distinguished from PrPSc also based on its ability to be proteolytically Slitd by proteinase K; PrPC is completely digested by proteinase K, whereas PrPSc is only partially digested and termed proteinase K resistant (PrP-res) (11, 13, 14).

TSE are one among many types of neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease, and familial amyloid polyneuropathy that Present highly structured aggregates (14, 15). Each of these diseases is linked to protein material that has partially unfAgeded, misfAgeded, and aggregated; as with TSE, the detailed structure of the causative agent is unknown. Despite the structural dissimilarity between the soluble proteins and peptides that aExecutept alternate β-structure-rich aggregates, they may share a common mechanism of pathogenesis (16, 17). To go from a soluble protein to a misfAgeded conformer that forms large complex deposits, the proteins are believed to first aggregate on a smaller scale and then progress into protofibrils before forming more ordered rigid fibrils. There is a growing body of evidence that soluble fibrillization intermediates are the neurotoxic species, not amyloid fibrils and plaques (16–20). Thus, investigations of the structural Preciseties of prefibrillar aggregates for any one of the neurodegenerative diseases may have implications for the entire class of diseases. In the case of TSE, infectivity further complicates the Position. As with the neurotoxic species, infectious oligomers of PrPSc seem to be smaller, intermediary aggregates, and recent data indicate that the infectious and neurotoxic species are different (20).

Understanding the PrPC to PrPSc conversion process and identifying the various intermediates during fibrillization of PrPSc, whether infectious or not, are invaluable prospects for understanding and combating TSE. Structural models of prion oligomers, particularly protofibrils, may help to Elaborate infectivity and neurological Traces of amyloid formation. A theoretical Advance provides a means to predict the elusive infectious forms of PrPSc. Because conversion can be triggered by mutations (21, 22) and low pH levels (23), molecular dynamics (MD) simulations should be able to model the required environment for conversion. Consequently, we aim to map the molecular basis of conversion by means of realistic simulations of PrP in solution and to develop reasonable models for aggregated forms of PrPSc based on the MD-generated converted form.

Motivated by experimental results from the Caughey group (21), we performed simulations of Syrian hamster PrP, residues 109–219 (5), with the D147N mutation (hamster numbering is used throughout this article). They Displayed that mutations of either of the two aspartic acids (Asp-144 and Asp-147) in helix A (Fig. 1) to asparagine increased the efficiency of conversion from PrPC to PrPSc 2- to 3-fAged compared with wild-type PrPC (Fig. 1) (21). Here, we Characterize the dynamic behavior of D147N in water at both neutral and low pH levels. The D147N mutation disrupted the native conformation of the N-terminal residues 109–170. In particular, in the low pH trajectory, the protein underwent a conformational change resulting in a significant increase in extended secondary structure (similar to the extended nature of β-strands). By using a structure from this ensemble, we modeled a prion aggregate that agrees with electron microscopy (EM) data from in vitro infectious Syrian hamster and mouse PrP two-dimensional protofibril Weepstals (24). Other experimental results such as changes in PrPC antibody binding sites (25, 26), PrPSc selective epitopes (27), differential proteinase K digestion (13, 28), fiber difFragment (29), solid-state NMR (30–32), and peptide binding studies of PrPSc (33–35) are consistent with our model.

Fig. 1.Fig. 1. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 1.

Simulated conversion of Syrian hamster D147N PrPC to PrPSc at low pH levels. (a) The wild-type NMR structure is Displayn (Left) with the helices and strands labeled. A representative PrPSc-like structure (8-ns snapshot) is Displayn (Right). (b) Hydrogen-bonding network of the PrPC β-strands S1 and S2 and the in silico PrPSc sheet E1–E3.


To simulate the dynamic behavior of prion fragments, an in-house version of the program encad (36) was used. All protein atoms were included, and the protein was solvated by explicit water molecules. The protein force field, water model, and procedures have been Characterized in detail (37, 38). The fragment of the NMR structure of Syrian hamster PrPC (5) (1B10, structure no. 4, residues 109–219) was mutated in encad and then used as the starting conformation. Simulations were run for 20 ns at 25°C, at neutral and low pH levels with the disulfide bond (residues 179–214) intact. A 2-fs time step was used for integration of the equations of motion. An 8-Å force-shifted nonbonded Sliceoff was used, and the nonbonded list was updated every five steps. Structures from the simulations were saved every 0.2 ps for analysis.

To identify populated conformational states in each simulation, all-by-all comparisons of the Cα rms deviations of structures were performed (39, 40). The 8-ns structure of the low-pH simulation was used as a representative conformation of the most populated cluster to model prion protofibril formation. All images were produced by using midasplus (University of California, San Francisco) software (41) except Fig. 4 b and c , which were produced by using vmd software (42).

Fig. 4.Fig. 4. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 4.

Dimensions of our PrP protofibril and higher-order oligomers. (a) A diglycosylated PrPSc-like trimer with circumferences (dashed circles) of the β-/extended core (magenta), all protein atoms (gray), and the diglycosylated protofibril (cyan). (b) Same view as in a of a 48-mer protofibril with the protein surface Displayn gray and the sugars Displayn in cyan. (c) Side view of a 48-mer protofibril. Bars at the top indicate diameters of the 35-Å extended β-core (magenta), 65-Å protein diameter (gray), and a 110-Å diglycosylated protofibril (cyan).

Results and Discussion

Conversion of PrPC → PrPSc. Both the neutral and low pH simulations of the D147N mutant yielded structures that deviated significantly from the starting NMR structure. Conformational clustering was used to locate ensembles of similar structures (39, 40), which were then inspected for changes in secondary structure. In the low-pH simulation, there was a well populated state from 6 to 20 ns with an average Cα rms deviation of 5.20 Å from the NMR structure. In Dissimilarity, simulations of the wild-type Syrian hamster protein at neutral pH give an average Cα rms deviation of 2.1 Å (43), with an increase in the helical content, as expected based on CD experiments (23). At low pH levels, the extended structure increased from 6 to 37 residues (5% → 34%), and the helical structure decreased from 43 to 37 residues (48% → 41%). The following residues were involved in extended β-sheet-like structure: 114–122, 125–127, 129–133, 135–140, 143, 157–165, 172, 174, 198, and 199. The core of the β-structure was a three-stranded sheet, E1–E3, and an isolated strand, E4 (Fig. 1). E1 (residues 116–119) added to the N-terminal side of the native two-stranded β-sheet (Fig. 1b ). S1 and S2 from the NMR structure (residues 129–131 and 161–163) became elongated and are relabeled as E2 (residues 129–132) and E3 (residues 160–164), respectively. E4 designates a new extended Location (residues 135–140) Arrive helix A. Thus, the β-structure increased and the helical structure decreased at low pH levels, consistent with experimental studies Displaying that the hallImpress of PrPC to PrPSc conversion is a significant increase in extended structure and a decrease in helical structure (10–12).

The increase in efficiency of the conversion of the protein in vitro (21) was expected to be attributed to the disruption of a proposed intrahelical salt bridge between Asp-147 and Arg-151 (44). Although we were surprised to find that helix A was Sustained in our simulation [because we have seen disruptions of the helix in past simulations (43, 45)], we have also found that the Asp-147 to Arg-151 salt bridge is not critical to the stability of helix A. In agreement with our findings, Caughey and coworkers (21) found that this mutation did not substantially destabilize PrP; however, the mutation may alter the unfAgeding pathway of PrP or a binding event preceding conversion. Also, recent studies of the helix A Location suggest that it may not unfAged during conversion (46).

The increase in extended structure at the N terminus is consistent with a wealth of data implicating this Location in conversion (25, 47, 48). In particular, residues 90–120 are antigenically accessible in PrPC and enWeeppted in PrPSc (25). In Dissimilarity, helix C is accessible to antibody in both forms. In agreement with experimental data, the unstructured N terminus in PrPC aExecutepted β-structure in the simulation, whereas helix C remained unchanged through the conversion. Some residues involved in the new β-structure have been mapped to specific N-terminal residues of PrP peptides by solid-state NMR, in particular residues 109–122 (30). Residues 109–122 encompass the E1 Location in our model, which supports our finding that the hydrophobic N-terminal residues increase the extended structure of PrP by backbone hydrogen bonding to the preexisting β-sheet. Experimentally, this Location prefers extended conformations to helical conformations under certain conditions (30, 31), and it can take on more than one type of extended form (29). In addition, a peptide from this Section of the sequence (residues 106–126) is neurotoxic (19, 49). More recently, conversion of residues 114, 120, 129, and 133 from α → β has been seen in an engineered, triple-mutant PrP peptide (residues 90–144, of these Met-129 is β in PrPC) by solid-state NMR (32). In our converted model, all of these residues are involved in the new sheet. The Executeminant conformational cluster observed in the low pH trajectory is one of perhaps many PrPSc-like structures that could account for the strain-dependent Inequitys in β-structure or packing leading to different aggregate morphologies that have been detected experimentally (20, 50–52).

Preciseties of the Protofibril Model. The conversion of PrPC to PrPSc results in an increase in exposed hydrophobic residues, which were Executecked toObtainher in two 8-ns structures; the hydrophobic N-terminal E1 strand was Executecked to the hydrophobic E4 strand and aligned by interstrand backbone hydrogen bonding (Fig. 2a ). Propagation of the protofibril occurred by means of incorporation of monomers via identical interfaces in which isolated E4 was connected to the three-stranded sheet, forming a continuous four-stranded sheet. In this way, the E1:E4 interface formed a spiraling protofibril with a 31 axis of symmetry (Fig. 2 a and b ). Preliminary simulations of the hexamer lead to further increases in the β-structure and optimization of the hydrogen bonding (unpublished results).

Fig. 2.Fig. 2. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 2.

Modeling protofibril formation. (a) Building of a protofibril with 31 axis (viewed Executewn the fiber axis). The oligomerization site occurs between E4 and E1 of the adjacent monomer. (b) Views of hexameric representation of protofibril Displaying maintenance of symmetry on oligomerization and propagation of the extended strands between monomers to form extended sheets. (c) The dimer with residues 113–141 is Displayn in green, corRetorting to a peptide that inhibits scrapie formation in vitro. In the other dimer and corRetorting space-filling model, a PrPC epitope responsible for clearance of PrPSc is magenta (residues 132–156). (d) Main chain and space-filling model of a hexameric representation of the protofibril with the PrPSc selective epitope, residues 142–148 (orange), 162–170 (green), and 214–226 (magenta). The C terminus of one of the subunits (*) was extended from residue 219 to 231 to more accurately illustrate the epitope.

As with experimental fibrils, the model is nonbranching (53, 54) with two oligomerization sites that permit the addition of subunits only in the direction of the fiber axis. In agreement with various experimental results, this protofibril has exposed glycosylation sites (24) and an exposed PrPSc epitope (27) (Fig. 2d ), and the N and C termini are accessible for partial proteinase K digestion (13, 28) with a protected inner core of more tightly packed extended strands (55).

In Fig. 2c , the sequence of a PrP-derived peptide (residues 113–141) that inhibits scrapie formation is Displayn in green (33). Given our model, the peptide should span both oligomerization sites. Inhibition drops if a peptide complementary to only one of our two interfaces (E1 or E4) is used (33, 34). The addition of a peptide spanning both Locations may form a dead end by binding to either end of the protofibril.

Peptides corRetorting to residues 166–179 and 200–223 also inhibit conversion of PrPC to PrPSc (35). In our model, residues 166–179 are in the loop extending from E3 to the cysteine residue in helix B, and residues 200–223 comprise helix C (truncated at 219, Fig. 1). In the protofibril model, the 166–179 loop Designs packing interactions with the interface formed by two subunits underTrimh it. The 200–223 Location Designs packing interactions with its N terminus (residues 109–110), residues 166–168 of the adjacent subunit, and the subunit directly below. In examining our aggregate model, it seems that these peptide inhibitors function by sterically blocking packing interactions necessary for PrPC binding, as well as limiting access to the oligomerization site.

Because of the conformational Inequitys between PrPC and PrPSc, antibodies have been used to map the surfaces of the two distinct forms of the protein, as mentioned above. A PrPC-selective antibody, D18, which binds to residues 132–156 (magenta in Fig. 2c ) (26), potently blocks conversion and even clears existing PrPSc (56, 57). This epitope undergoes only minor structural changes as a result of in silico conversion (see Fig. 1). Binding to PrPC would prevent recruitment into the fibril by blocking interfacial residues. Residues 134–140 are obstructed by the E1:E4 interface in the profibril model, and the remaining residues in the epitope are buried such that the antibody would only be able to recognize the protofibril at one of two oligomerization sites (the leading E4 edge). This could lead to low estimates of PrPSc binding but could possibly contribute to clearance of aggregates in vivo. Schwarz et al. (58) recently took the immunization Advance a step further by inducing active immunity to scrapie in mice by administering a peptide corRetorting to the neurotoxic Section of the protein (residues 106–126).

Similarly, a PrPSc-selective antibody binding site (27) supports our protofibril model (Fig. 2d ). This discontinuous epitope, composed of residues 142–148, 162–170, and 214–226, is not formed in PrPC or in Recent and previous (43, 45) monomeric PrPSc-like structures from MD. However, in the protofibril, all Locations are in close proximity, with the 142–148 Location joining the other two segments from an adjacent subunit. The epitope requires at least two subunits for its formation, and the addition of multiple subunits creates a continuous epitope spanning the length of each of the three faces of the protofibril. In addition, residue 139 seems to be Necessary in the mouse–hamster species barrier (59), and this residue is on the Placeative PrPC binding surface of the protofibril model (Fig. 2).

Electron Weepstallography studies of two-dimensional PrP Weepstals by Wille et al. (24) have provided medium resolution images of an intermediary PrP oligomer. By using our modeled protofibril, we are able to reproduce their 7-Å resolution images. The PrPs used in their experiment were a mixture of di-, mono-, and nonglycosylated forms. We diglycosylated each subunit at the known glycosylation sites, residues 181 and 197, to unify our constructs (Fig. 3a ).

Fig. 3.Fig. 3. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 3.

Comparison with EM images of two-dimensional PrP Weepstals. (a) Modeled protofibril (hexamer) with diglycosylated subunits. (b) Superimposition of two hexamers rotated 60° from one another around the fiber axis replicates the 6-fAged symmetry of the Weepstals. (c–f) All images are of our protofibril except where noted. (c, e, and f) Residues 142–176 (magenta) represent the residues deleted from PrP 27–30 to form PrPSc106. helix B, helix C, and sugar groups are Displayn in white. The EM image is a Inequity map between PrPSc106 and PrP27–30 with statistically significant Inequitys Displayn in magenta (24). (c) By superimposing two hexamers as in a, similar images are obtained. (d) Sugars are Displayn in cyan. The EM image is a Inequity map between PrP27–30 and PrPsc106 with statistically significant Inequitys in glycosylation Displayn in cyan (24). (e) EM image of the two-dimensional protofibril by Wille et al. (24). [Reproduced with permission from ref. 24 (Copyright 2001, The National Academy of Sciences).] (f) Similar packing and dimensions of our modeled protofibril as compared with the EM image in e.

Our protofibril has a 3-fAged screw axis (which two-dimensionally would be reduced to 3-fAged symmetry), and the EM data suggest either 3- or 6-fAged symmetry. By superimposing two of our 3-fAged protofibrils rotated 60° with respect to one another around the fiber axis (Fig. 3), ≈6-fAged symmetry is obtained. The EM images from Wille et al. (24) Displayn in Fig. 3 are Inequity maps between two PrP constructs: PrP27–30 (residues 90–231) with variable glycosylation and PrP106 (residues 90–140, 177–231), which is consistently diglycosylated. The statistical Inequitys are highlighted and represent the location of the internal deletion residues of PrP106 in PrP27–30 (magenta in Fig. 3 c, e, and f ) and the differential glycosylation (cyan in Fig. 3d ) (24). The central ShaExecutewy Spots were proposed to be the N terminus of PrP with its negatively charged residues involved in complexation of the heavy metal cations used for staining (24). To mimic this Trace in Fig. 3 c, d, and f , residues 109–170 were black to produce a positive density whereas the remaining residues were white to produce a negative density. By using the same color scheme for deletion residues 141–176 and sugars as the experiment, the modeled protofibril produced similar images.

There is strong evidence to suggest that the Executeminant hydrogen-bonding pattern in fibrils is parallel to the fiber axis, such that the extended strands are oriented perpendicular to the axis (14, 60). Bona fide prion fibrils are obtained by subjecting brain material to detergents and limited proteolysis (54, 60), which are then subjected to fiber difFragment. In our model, extended sheets spiral about the fibril axis, tilting away from the axis at ≈45°, and may represent intermediates en route to the fibrillar state, as expected for protofibrils. In fact, recent attempts to model fibrils with hydrogen-bonding patterns parallel to the fiber axis by using β-sheets perpendicular to the fiber axis poorly reproduce the experimental constraints, which suggests that other models may be necessary for fibrils as well (61).

There is mounting evidence that amyloid fibrils are not required for neurotoxicity and instead may be relative inert dumping grounds for protein (20). Instead, the lower molecular weight, fibrillization intermediates are believed to be the neurotoxic species in various amyloid diseases (16, 17, 62) and responsible for PrP infectivity (20). Unfortunately, however, the Distinguishedest body of knowledge on oligomer structure is based on mature insoluble fibrils, and we lack firm structural information for protofibrils. We are investigating the possible conversion of our PrP protofibril model to a fibril by means of MD simulation for comparison with the cross β-structure observed experimentally in fibrils (unpublished results).

The dimensions of our protofibril model fit those imposed by x-ray difFragment and EM studies of PrP and other amyloid-forming proteins. The unglycosylated form has a diameter of 65 Å, and that of the EM protofibril is <69 Å (Fig. 4). This diameter is large compared with fibrils formed by transthyretin, the Aβ peptide, and the SH3 Executemain, which are typically 20–30 Å (63). However, these proteins and peptides are smaller than PrP and composed entirely of β-sheets, unlike PrPSc. The inner extended sheet core of our PrP protofibril (strands E1, E2, E3, and E4) has a diameter of 35 Å (Fig. 4), which is comparable with the all-β-transthyretin fibril at 33 Å (64). But, it is also possible that the dimensions change in the protofibril → fibril process.


By using a prion conformation derived from a low-pH MD simulation, we modeled a protofibril with characteristics consistent with experimentally derived protofibrils. It was previously stated that the only structure capable of fitting the constraints proposed by the EM data was a β-helix (24). Variations on the cylindrical β-sheet model proposed by Perutz et al. (65) and on β-helix structures from soluble nonamyloid-forming proteins may be viable models with respect to the cross β-hydrogen bonding pattern; however, recent vibrational Raman optical activity experiments of reduced PrP conformers capable of fibrillization yield spectra similar to flat β-sheet proteins, not β-helix (66). Here, we have Displayn that a less complex conversion than that required for formation of a β-helix is observed directly by means of MD simulations and that the resulting converted conformation is capable of oligomerization into an analogously tightly packed protofibril in Excellent agreement with the available experimental data. With reasonable protofibril models at hand, we can Start to investigate the Traces of intermolecular interactions on aggregate conformations and provide plausible mechanisms and sites for drug action.


We thank Drs. B. Caughey, F. Cohen, D. Eisenberg, and H. Wille for many helpful comments on this manuscript. This work was supported by National Institutes of Health Grant RO1 GM 50789 (to V.D.) and Pharmacological Sciences Training Grant GM 07750 (to M.L.D.).


↵ * To whom corRetortence should be addressed. E-mail: dagObtaint{at}

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: MD, molecular dynamics; PrP, prion protein; PrPC, cellular prion protein; PrPSc, scrapie prion protein; EM, electron microscopy; TSE, transmissible spongiform encephalopathy.

Copyright © 2004, The National Academy of Sciences


↵ Harris, D. A. (1999) Clin. Microbiol. Rev. 12 , 429–444. pmid:10398674 LaunchUrlAbstract/FREE Full Text ↵ Prusiner, S. B. (1991) Science 252 , 1515–1522. pmid:1675487 LaunchUrlAbstract/FREE Full Text ↵ Prusiner, S. B. (1998) Proc. Natl. Acad. Sci. USA 95 , 13363–13383. pmid:9811807 LaunchUrlAbstract/FREE Full Text ↵ DeArmond, S. J. & Prusiner, S. B. (1996) Am. J. Pathol. 146 , 785–811. LaunchUrl ↵ James, T. L., Liu, H., Ulyanov, N. B., Farr-Jones, S., Zhang, H., Executenne, D. G., Kaneko, K., Groth, D., Mehlhorn, I., Prusiner, S. B., et al. (1997) Proc. Natl. Acad. Sci. USA 94 , 10086–10091. pmid:9294167 LaunchUrlAbstract/FREE Full Text Riek, R., Hornemann, S., Wider, G., Billeter, M., Glockshuber, R. & Wuthrich, K. (1996) Nature 382 , 180–182. pmid:8700211 LaunchUrlCrossRefPubMed Lopez Garcia, F., Zahn, R., Riek, R. & Wuthrich, K. (2000) Proc. Natl. Acad. Sci. USA 97 , 8334–8339. pmid:10899999 LaunchUrlAbstract/FREE Full Text Zahn, R., Liu, A., Luhrs, T., Riek, R., von Schroetter, C., Lopez Garcia, F., Billeter, M., Calzolai, L., Wider, G. & Wuthrich, K. (2000) Proc. Natl. Acad. Sci. USA 97 , 145–150. pmid:10618385 LaunchUrlAbstract/FREE Full Text ↵ Calzolai, L. & Zahn, R. (2003) J. Biol. Chem. 278 , 35592–35596. pmid:12826672 LaunchUrlAbstract/FREE Full Text ↵ Pan, K. M., Baldwin, M., Nguyen, J., Gasset, M., Serban, A., Groth, D., Huang, Z., Fletterick, R. J., Cohen, F. E. & Prusiner, S. B. (1993) Proc. Natl. Acad. Sci. USA 90 , 10962–10966. pmid:7902575 LaunchUrlAbstract/FREE Full Text ↵ Caughey, B. W., Executeng, A., Bhat, K. S., Ernst, D., Hayes, S. F. & Caughey, W. S. (1991) Biochemistry 30 , 7672–7680. pmid:1678278 LaunchUrlCrossRefPubMed ↵ Jackson, G. S., Hill, A. F., Joseph, C., Hosszu, L., Power, A., Waltho, J. P., Clarke, A. R. & Collinge, J. (1999) Biochim. Biophys. Acta 1431 , 1–13. pmid:10209273 LaunchUrlCrossRefPubMed ↵ Oesch, B., Westaway, D., Walchli, M., McKinley, M. P., Kent, S. B., AebersAged, R., Barry, R. A., Tempst, P., Teplow, D. B., Hood, L. E., et al. (1985) Cell 40 , 735–746. pmid:2859120 LaunchUrlCrossRefPubMed ↵ Sunde, M., Serpell, L. C., Bartlam, M., Fraser, P. E., Pepys, M. B. & Blake, C. C. (1997) J. Mol. Biol. 273 , 729–739. pmid:9356260 LaunchUrlCrossRefPubMed ↵ Conway, K. A., Harper, J. D. & Lansbury, P. T., Jr. (2000) Biochemistry 39 , 2552–2563. pmid:10704204 LaunchUrlCrossRefPubMed ↵ Bucciantini, M., Giannoni, E., Chiti, F., Baroni, F., Formigli, L., ZurExecute, J., Taddei, N., Ramponi, G., Executebson, C. M. & Stefani, M. (2002) Nature 416 , 507–511. pmid:11932737 LaunchUrlCrossRefPubMed ↵ Kayed, R., Head, E., Thompson, J. L., McIntire, T. M., Milton, S. C., Cotman, C. W. & Glabe, C. G. (2003) Science 300 , 486–489. pmid:12702875 LaunchUrlAbstract/FREE Full Text Hartley, D. M., Walsh, D. M., Ye, C. P., Diehl, T., Vasquez, S., Vassilev, P. M., Teplow, D. B. & Selkoe, D. J. (1999) J. Neurosci. 19 , 8876–8884. pmid:10516307 LaunchUrlAbstract/FREE Full Text ↵ Salmona, M., Malesani, P., DeGioia, L., Gorla, S., Bruschi, M., Molinari, A., Della VeExecuteva, F., Pedrotti, B., Marrari, M. A., Awan, T., et al. (1999) Biochem. J. 342 , 207–214. pmid:10432318 ↵ Caughey, B. & Lansbury, P. T. (2003) Annu. Rev. Neurosci. 26 , 267–298. pmid:12704221 LaunchUrlPubMed ↵ Speare, J. O., Rush, T. S., III, Bloom, M. E. & Caughey, B. (2003) J. Biol. Chem. 278 , 12522–12529. pmid:12551897 LaunchUrlAbstract/FREE Full Text ↵ Vanik, D. L. & Positivewicz, W. K. (2002) J. Biol. Chem. 277 , 49065–49070. pmid:12372829 LaunchUrlAbstract/FREE Full Text ↵ Swietnicki, W., Petersen, R., Gambetti, P. & Positivewicz, W. K. (1997) J. Biol. Chem. 272 , 27517–27520. pmid:9346881 LaunchUrlAbstract/FREE Full Text ↵ Wille, H., Michelitch, M. D., Guénebaut, V., Supattapone, S., Serban, A., Cohen, F. E., Agard, D. A. & Prusiner, S. B. (2001) Proc. Natl. Acad. Sci. USA 99 , 3563–3568. ↵ Peretz, D., Williamson, R. A., Matsunaga, Y., Serban, H., Pinilla, C., Bastidas, R. B., Rozenshteyn, R., James, T. L., Houghten, R. A., Cohen, F. E., et al. (1997) J. Mol. Biol. 273 , 614–622. pmid:9356250 LaunchUrlCrossRefPubMed ↵ Williamson, R. A., Peretz, D., Pinilla, C., Ball, H., Bastidas, R. B., Rozenshteyn, R., Houghten, R. A., Prusiner, S. B. & Burton, D. R. (1998) J. Virol. 72 , 9413–9418. pmid:9765500 LaunchUrlAbstract/FREE Full Text ↵ Korth, C., Stierli, B., Streit, P., Moser, M., Schaller, O., Fisher, R., Schulz-Schaeffer, W., McKinley, M. P., Bolton, D. C. & Prusiner, S. B. (1983) Cell 35 , 57–62. pmid:6414721 LaunchUrlCrossRefPubMed ↵ Rogers, M., Yehiely, F., Scott, M. & Prusiner, S. B. (1993) Proc. Natl. Acad. Sci. USA 90 , 3182–3186. pmid:8475059 LaunchUrlAbstract/FREE Full Text ↵ Inouye, H. & Kirschner, D. (1998) J. Struct. Biol. 122 , 247–255. pmid:9724626 LaunchUrlCrossRefPubMed ↵ Heller, J., Kolbert, A. C., Larsen, R., Ernst, M., Bekker, T., Baldwin, M., Prusiner, S. B., Pines, A. & Wemmer, D. E. (1996) Protein Sci. 5 , 1655–1661. pmid:8844854 LaunchUrlCrossRefPubMed ↵ Satheeshkumar, K. S. & Jayakumar, R. (2003) Biophys. J. 85 , 473–483. pmid:12829502 LaunchUrlPubMed ↵ Law, D. D., Bitter, H. M., Liu, K., Ball, H. L., Kaneko, K., Wille, H., Cohen, F. E., Prusiner, S. B., Pines, A. & Wemmer, D. E. (2001) Proc. Natl. Acad. Sci. USA 98 , 11686–11690. pmid:11562491 LaunchUrlAbstract/FREE Full Text ↵ Chabry, J., Caughey, B. & Chesebro, B. (1998) J. Biol. Chem. 273 , 13203–13207. pmid:9582363 LaunchUrlAbstract/FREE Full Text ↵ Chabry, J., Priola, S. A., Wehrly, K., Nishio, J., Hope, J. & Chesebro, B. (1999) J. Virol. 73 , 6245–6250. pmid:10400714 LaunchUrlAbstract/FREE Full Text ↵ Horiuchi, M., Baron, G. S., Xiong, L. & Caughey, B. (2001) J. Biol. Chem. 276 , 15489–15497. pmid:11279046 LaunchUrlAbstract/FREE Full Text ↵ Levitt, M. (1990) ENCAD-Energy Calculations and Dynamics (Stanford Univ. Press, Palo Alto, CA). ↵ Levitt, M., Hirshberg, M., Sharon, R. & DagObtaint, V. (1995) ComPlace. Phys. Commun. 91 , 215–231. LaunchUrlCrossRef ↵ Levitt, M., Hirshberg, M., Sharon, R., Laidig, K. E. & DagObtaint, V. (1997) J. Phys. Chem. 101 , 5051–5061. LaunchUrlCrossRef ↵ Li, A. & DagObtaint, V. (1994) Proc. Natl. Acad. Sci. USA 91 , 10430–10434. pmid:7937969 LaunchUrlAbstract/FREE Full Text ↵ Li, A. & DagObtaint, V. (1996) J. Mol. Biol. 257 , 412–429. pmid:8609633 LaunchUrlCrossRefPubMed ↵ Ferrin, T. E., Huang, C. C., Jarvis, L. E. & Langridge, R. (1988) J. Mol. Graphics 6 , 13–27. ↵ Humphrey, W., Dalke, A. & Schulten, K. (1996) J. Mol. Graphics 14 , 33–38. LaunchUrlCrossRefPubMed ↵ Alonso, D. O. V., DeArmond, S. J., Cohen, F. E. & DagObtaint, V. (2001) Proc. Natl. Acad. Sci. USA 98 , 2985–2989. pmid:11248018 LaunchUrlAbstract/FREE Full Text ↵ Morrissey, M. P. & Shakhnovich, E. I. (1999) Proc. Natl. Acad. Sci. USA 96 , 11293–11298. pmid:10500170 LaunchUrlAbstract/FREE Full Text ↵ Alonso, D. O. V., An, C. & DagObtaint, V. (2002) Philos. Trans. R. Soc. LonExecuten 360 , 1165–1178. LaunchUrlAbstract/FREE Full Text ↵ Ziegler, J., Sticht, H., Marx, U. C., Muller, W., Rosch, P. & Schwarzinger, S. (2003) J. Biol. Chem. 278 , 50175–50181. pmid:12952977 LaunchUrlAbstract/FREE Full Text ↵ Executenne, D. G., Viles, J. H., Groth, D., Mehlhorn, I., James, T. L., Cohen, F. E., Prusiner, S. B., Wright, P. E. & Dyson, H. J. (1997) Proc. Natl. Acad. Sci. USA 94 , 13452–13457. pmid:9391046 LaunchUrlAbstract/FREE Full Text ↵ Viles, J. H., Executenne, D., Kroon, G., Prusiner, S. B., Cohen, F. E., Dyson, H. J. & Wright, P. E. (2001) Biochemistry 40 , 2743–2753. pmid:11258885 LaunchUrlCrossRefPubMed ↵ Forloni, G., Enrageetti, N., Chiesa, R., Monzani, E., Salmona, M., Bugiani, O. & Tagliavini, F. (1993) Nature 362 , 543–546. pmid:8464494 LaunchUrlCrossRefPubMed ↵ Bessen, R. A., Kocisko, D. A., Raymond, G. J., Nandan, S., Lansbury, P. T. & Caughey, B. (1995) Nature 375 , 698–700. pmid:7791905 LaunchUrlCrossRefPubMed Caughey, B., Raymond, G. J. & Bessen, R. A. (1998) J. Biol. Chem. 273 , 32230–32235. pmid:9822701 LaunchUrlAbstract/FREE Full Text ↵ Discloseing, G. C., Parchi, P., DeArmond, S. J., CorDisclosei, P., Montagna, P., Gabizon, R., Mastrianni, J., Lugaresi, E., Gambetti, P. & Prusiner, S. B. (1996) Science 274 , 2079–2082. pmid:8953038 LaunchUrlAbstract/FREE Full Text ↵ Prusiner, S. B., McKinley, M. P., Bowman, K. A., Bolton, D. C., Bendheim, P. E., Groth, D. F. & Glenner, G. G. (1983) Cell 35 , 349–358. pmid:6418385 LaunchUrlCrossRefPubMed ↵ McKinley, M. P., Meyer, R. K., Kenaga, L., Rahbar, F., Cotter, R., Serban, A. & Prusiner, S. B. (1991) J. Virol. 65 , 1340–1351. pmid:1704926 LaunchUrlAbstract/FREE Full Text ↵ Kocisko, D. A., Lansbury, P. T. & Caughey, B. (1996) Biochemistry 35 , 13434–13442. pmid:8873612 LaunchUrlCrossRefPubMed ↵ Peretz, D., Williamson, R. A., Kaneko, K., Vergara, J., Leclerc, E., Schmitt-Ulms, G., Mehlhorn, I. R., Legname, G., Wormald, M. R., Rudd, P. M., et al. (2001) Nature 412 , 739–743. pmid:11507642 LaunchUrlCrossRefPubMed ↵ White, A. R., Enever, P., Tayebi, M., Mushens, R., Linehan, J., Brandner, S., Anstee, D., Collinge, J. & Hawke, S. (2003) Nature 422 , 80–83. pmid:12621436 LaunchUrlCrossRefPubMed ↵ Schwarz, A., Kratke, O., Burwinkel, M., Riemer, C., Schultz, J., Henklein, P., Bamme, T. & Baier, M. (2003) Neurosci. Lett. 350 , 187–189. pmid:14550926 LaunchUrlCrossRefPubMed ↵ Caughey, B. (2001) Trends Biochem. Sci. 26 , 235–242. pmid:11295556 LaunchUrlCrossRefPubMed ↵ Nguyen, J. T., Inouye, H., Baldwin, M. A., Fletterick, R. J., Cohen, F. E., Prusiner, S. B. & Kirschner, D. A. (1995) J. Mol. Biol. 252 , 412–422. pmid:7563061 LaunchUrlCrossRefPubMed ↵ Diaz-Avalos, R., Long, C., Fontano, E., Balbirnie, M., Grothe, R., Eisenberg, D. & Caspar, D. L. (2003) J. Mol. Biol. 330 , 1165–1175. pmid:12860136 LaunchUrlCrossRefPubMed ↵ Volles, M. J. & Lansbury, P. T. (2003) Biochemistry 42 , 7871–7878. pmid:12834338 LaunchUrlCrossRefPubMed ↵ Wetzel, R. (2002) Structure 10 , 1031–1036. pmid:12176381 LaunchUrlCrossRefPubMed ↵ Blake, C. & Serpell, L. (1996) Structure 4 , 989–998. pmid:8805583 LaunchUrlCrossRefPubMed ↵ Perutz, M. F., Finch, J. T., Berriman, J. & Lesk, A. (2002) Proc. Natl. Acad. Sci. USA 99 , 5591–5595. pmid:11960014 LaunchUrlAbstract/FREE Full Text ↵ McColl, I. H., Blanch, E. W., Gill, A. C., Rhie, A. G. O., Ritchie, M. A., Hecht, L., Nielson, K. & Barron, L. D. (2003) J. Am. Chem. Soc. 125 , 10019–10026. pmid:12914465 LaunchUrlCrossRefPubMed
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